MAX™ VAGINAL PANEL 443712 For In Vitro Diagnostic Use P0223(08) For use with the BD MAX™ System 2020-05 English INDICATIONS FOR USE The BD MAX™ Vaginal Panel performed on the BD MAX System is an automated qualitative in vitro diagnostic test for the direct detection of DNA targets from bacteria associated with bacterial vaginosis (qualitative results reported based on detection and quantitation of targeted organism markers), Candida species associated with vulvovaginal candidiasis, and Trichomonas vaginalis from vaginal swabs in patients who are symptomatic for vaginitis/vaginosis. The test utilizes real-time polymerase chain reaction (PCR) for the amplification of specific DNA targets and utilizes fluorogenic target-specific hybridization probes to detect and differentiate DNA from: • Bacterial vaginosis markers (Individual markers not reported) Lactobacillus spp. (L. crispatus and L. jensenii) Gardnerella vaginalis Atopobium vaginae Bacterial Vaginosis Associated Bacteria-2 (BVAB-2) Megasphaera-1 • Candida spp. (C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis) • Candida glabrata • Candida krusei • Trichomonas vaginalis The BD MAX Vaginal Panel is intended to aid in the diagnosis of vaginal infections in women with a clinical presentation consistent with bacterial vaginosis, vulvovaginal candidiasis and trichomoniasis. SUMMARY AND EXPLANATION OF THE PROCEDURE Vaginitis, one of the most common gynecological problems in clinical medicine, accounts for millions of office visits each year. 1,2 The three main infectious causes of vaginitis are bacterial vaginosis (BV), yeast vaginitis (candidiasis) and T. vaginalis vaginitis (trichomoniasis). 2 Bacterial vaginosis is the most common infectious vaginal complaint, and accounts for 40 to 50 percent of cases in women of childbearing age. 2 Hydrogen-peroxide producing Lactobacillus species are important members of the normal vaginal flora and are generally known to decrease in patients suffering from vaginosis. The decrease of these bacteria rather than their colonization of the vagina is a recognized marker of vaginosis. 3,4 In opposition to this decrease in Lactobacillus species; there is an increase in well-known species of anaerobes such as Atopobium vaginae and Gardnerella vaginalis. 4 Colonization also occurs with non-cultivable anaerobe organisms such as BVAB-2 and Megasphaera-1. 3 Additional microorganisms including Prevotella, Lachnospira, Sneathia, Mobiluncus, Mycoplasma hominis, and Ureaplasma spp. have also been found in women with bacterial vaginosis. 5 However, these organisms are less associated with BV due to their relatively low prevalence, sensitivity and/or specificity. 3,6,7 Bacterial vaginosis is associated with a high incidence of endometritis and pelvic inflammatory disease following abortion and gynecologic procedures in the general population. Bacterial vaginosis has been also associated with late-term miscarriages, premature rupture of membranes, and preterm birth, and has been strongly linked with an increased risk of human immunodeficiency virus (HIV) and other sexually transmitted diseases. 8,9 Although it is known that asymptomatic women may have microbial flora consistent with bacterial vaginosis, treatment is generally not indicated by current guidelines for these patients. 1
Transcript
MAX™ VAGINAL PANEL
443712For In Vitro Diagnostic Use P0223(08)For use with the BD MAX™ System 2020-05 English
INDICATIONS FOR USEThe BD MAX™ Vaginal Panel performed on the BD MAX System is an automated qualitative in vitro diagnostic test for the direct detection of DNA targets from bacteria associated with bacterial vaginosis (qualitative results reported based on detection and quantitation of targeted organism markers), Candida species associated with vulvovaginal candidiasis, and Trichomonas vaginalis from vaginal swabs in patients who are symptomatic for vaginitis/vaginosis. The test utilizes real-time polymerase chain reaction (PCR) for the amplification of specific DNA targets and utilizes fluorogenic target-specific hybridization probes to detect and differentiate DNA from:
• Bacterial vaginosis markers (Individual markers not reported) Lactobacillus spp. (L. crispatus and L. jensenii) Gardnerella vaginalis Atopobium vaginae Bacterial Vaginosis Associated Bacteria-2 (BVAB-2) Megasphaera-1
• Candida spp. (C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis)• Candida glabrata• Candida krusei• Trichomonas vaginalis
The BD MAX Vaginal Panel is intended to aid in the diagnosis of vaginal infections in women with a clinical presentation consistent with bacterial vaginosis, vulvovaginal candidiasis and trichomoniasis.
SUMMARY AND EXPLANATION OF THE PROCEDUREVaginitis, one of the most common gynecological problems in clinical medicine, accounts for millions of office visits each year.1,2 The three main infectious causes of vaginitis are bacterial vaginosis (BV), yeast vaginitis (candidiasis) and T. vaginalis vaginitis (trichomoniasis).2 Bacterial vaginosis is the most common infectious vaginal complaint, and accounts for 40 to 50 percent of cases in women of childbearing age.2 Hydrogen-peroxide producing Lactobacillus species are important members of the normal vaginal flora and are generally known to decrease in patients suffering from vaginosis. The decrease of these bacteria rather than their colonization of the vagina is a recognized marker of vaginosis.3,4 In opposition to this decrease in Lactobacillus species; there is an increase in well-known species of anaerobes such as Atopobium vaginae and Gardnerella vaginalis.4 Colonization also occurs with non-cultivable anaerobe organisms such as BVAB-2 and Megasphaera-1.3 Additional microorganisms including Prevotella, Lachnospira, Sneathia, Mobiluncus, Mycoplasma hominis, and Ureaplasma spp. have also been found in women with bacterial vaginosis.5 However, these organisms are less associated with BV due to their relatively low prevalence, sensitivity and/or specificity.3,6,7 Bacterial vaginosis is associated with a high incidence of endometritis and pelvic inflammatory disease following abortion and gynecologic procedures in the general population. Bacterial vaginosis has been also associated with late-term miscarriages, premature rupture of membranes, and preterm birth, and has been strongly linked with an increased risk of human immunodeficiency virus (HIV) and other sexually transmitted diseases.8,9 Although it is known that asymptomatic women may have microbial flora consistent with bacterial vaginosis, treatment is generally not indicated by current guidelines for these patients.
1
2
Seventeen (17) to 39 percent of women symptomatic for vaginitis are diagnosed with vulvovaginal candidiasis.9 Multiple epidemiologic studies have indicated that Candida albicans is responsible for 65 to 90% of vulvovaginal candidiasis cases10–15 and that non-Candida albicans species may be responsible for up to 30% of vulvovaginal candidiasis episodes.13,14,15 Among these species, the most frequently reported in literature are C. glabrata, C. parapsilosis and C. tropicalis. C. glabrata and C. krusei are the main Candida species resistant to fluconazole-based antifungal therapy.8,16,17 Vulvovaginal candidiasis is usually associated with high Candida loads but symptoms may be associated with lower loads in women predisposed by factors altering the vaginal milieu.18,19 Complications of vulvovaginal candidiasis are rare. Chorioamnionitis in pregnancy and vulvar vestibulitis syndrome have been reported.2 Women may also be colonized with Candida spp. and be asymptomatic. Trichomoniasis, caused by Trichomonas vaginalis, is one of the most common sexually transmitted infections worldwide with over 170 million cases per year.20 Four (4) to 35% of women symptomatic for vaginitis are diagnosed with trichomoniasis.9 This disease is associated with several adverse health outcomes, such as preterm birth, delivery of a low-birth weight infant, and facilitation of sexual transmission of Human Immunodeficiency Virus (HIV).1
Physicians traditionally diagnose vaginitis using the combination of symptoms, physical examination, pH of vaginal fluid, microscopy, and the whiff test. When combined, these tests display a sensitivity and specificity of 81 and 70% respectively for bacterial vaginosis; 84 and 85% for vulvovaginal candidiasis; and 85 and 100% for trichomoniasis when compared with a molecular assay.9, 21
The BD MAX Vaginal Panel is designed for use with the BD MAX UVE Specimen Collection kit. Samples are transported to the testing laboratory in BD MAX UVE Sample Buffer Tubes. The Sample Buffer Tubes are vortexed to release cells from the swab into the buffer. The Sample Buffer Tubes, Unitized Reagent Strips and PCR Cartridges are loaded on the BD MAX System. No further operator intervention is necessary and the following automated procedures occur. The cells are lysed, and DNA is extracted, captured and concentrated on magnetic beads. After an elution step, DNA is added to reagents containing specific primers and probes used to amplify and detect the genetic targets, if present. Upon amplification, signal detection and interpretation are performed automatically by the BD MAX System using real-time PCR. Each Extraction Tube includes a Sample Processing Control, which monitors the integrity of the reagents as well as the process steps involved in DNA extraction, amplification and detection, and checks for the presence of potential assay inhibitors.
PRINCIPLES OF THE PROCEDUREA combination of lytic and extraction reagents is used to perform cell lysis and DNA extraction. Nucleic acids released from the target organisms are captured on magnetic affinity beads. The beads, together with the bound nucleic acids, are washed and the nucleic acids are eluted by a combination of heat and pH variation. Eluted DNA is neutralized and transferred to the Master Mix Tubes to rehydrate the PCR reagents. After reconstitution, the BD MAX System dispenses a fixed volume of PCR-ready solution containing extracted nucleic acids into the PCR Cartridge. Microvalves in the cartridge are sealed by the system prior to initiating PCR in order to contain the amplification mixture and thus prevent evaporation and contamination. The amplified DNA targets are detected using hydrolysis (TaqMan®) probes, labeled at one end with a fluorescent reporter dye (fluorophore), and at the other end, with a quencher moiety. Probes labeled with different fluorophores are used to detect the target analytes in different optical channels of the BD MAX System. When the probes are in their native state, the fluorescence of the fluorophore is quenched due to its proximity to the quencher. However, in the presence of target DNA, the probes hybridize to their complementary sequences and are hydrolyzed by the 5’–3’ exonuclease activity of the DNA polymerase as it synthesizes the nascent strand along the DNA template. As a result, the fluorophores are separated from the quencher molecules and fluorescence is emitted. The amount of fluorescence detected in the optical channels used for the BD MAX Vaginal Panel is directly proportional to the quantity of the corresponding probe that is hydrolyzed. The BD MAX System monitors these signals at each cycle of the PCR and interprets the data at the end of the reaction to provide qualitative test results for each analyte of the vaginitis targets (i.e., positive or negative) and positive or negative BV results are obtained from the combination of bacterial vaginosis marker signals.
EQUIPMENT AND MATERIALS REQUIRED BUT NOT PROVIDED• BD MAX™ System (BD Cat. No. 441916)• BD MAX™ UVE Specimen Collection kit (BD Cat. No. 443376)• BD MAX™ PCR Cartridges (BD Cat. No. 437519)• BD MAX™ UVE Sample Buffer Tubes (BD Cat. No. 443420)• VWR Multi-Tube Vortexer (VWR Cat. No. 58816-115)• NALGENE™ Cryogenic Vial Holder (VWR Cat. No. 66008-783)• Suggested media for cultivation of external controls: Sabouraud Dextrose Agar (Deep Fill), (for example BD Cat. No. 221180 or
221278) or Sabouraud Dextrose Agar (Deep Fill), Emmons (for example BD Cat. No. 221867 or 221849), Columbia AnaerobicSheep Blood Agar (for example BD Cat. No. 221928).
• Disposable gloves, powderless
WARNINGS AND PRECAUTIONS• The BD MAX Vaginal Panel is for in vitro diagnostic use.• If patient-collected vaginal swabs are not directly transferred into a BD MAX UVE Sample Buffer Tube, they must be transferred
within 2 hours of collection when kept at 2–30 °C.• Do not pre-warm samples prior to using the BD MAX Vaginal Panel.• Do not use expired reagents and/or materials.• Do not use the kit if the label that seals the outer box is broken upon arrival.• Do not use reagents if the protective pouches are open or broken upon arrival.• Do not use reagents if desiccant is not present or is broken inside reagent pouches.• Do not remove desiccant from reagent pouches.• Close protective pouches of reagents promptly with the zip seal after each use. Remove any excess air in the pouches prior
to sealing.• Protect reagents against heat and humidity. Prolonged exposure to humidity may affect product performance.• Do not use reagents if the foil has been broken or damaged.• Do not mix reagents from different pouches and/or kits and/or lots.• Do not interchange or reuse caps, as contamination may occur and compromise test results.• Check Unitized Reagent Strips for proper liquid fills (ensure that the liquids are at the bottom of the tubes) (refer to Figure 2).• Check Unitized Reagent Strips to ensure that all pipette tips are present (refer to Figure 2).• Proceed with caution when using chemical solutions as Master Mix and Extraction Tube, barcode readability may be altered.• Good laboratory technique is essential to the proper performance of this assay. Due to the high analytical sensitivity of this test,
extreme care should be taken to preserve the purity of all materials and reagents.• In cases where other PCR tests are conducted in the same general area of the laboratory, care must be taken to ensure that
the BD MAX Vaginal Panel, any additional reagents required for testing, and the BD MAX System are not contaminated. Avoidmicrobial and deoxyribonuclease (DNase) contamination of reagents at all times. Gloves must be changed before manipulatingreagents and cartridges.
• To avoid contamination of the environment by amplicons, do not break apart the BD MAX PCR Cartridges after use. The sealsof the BD MAX PCR Cartridges are designed to prevent contamination.
• The laboratory should routinely perform environmental monitoring to minimize the risk of cross-contamination.• Performing the BD MAX Vaginal Panel outside the recommended time ranges can produce invalid results. Assays not
performed within the specified time ranges should be repeated with a new specimen.• Additional controls may be tested according to guidelines or requirements of local, state, provincial and/or federal regulations or
accrediting organizations.• Always handle specimens as if they are infectious and in accordance with safe laboratory procedures such as those described
in the CLSI Document M2922 and in Biosafety in Microbiological and Biomedical Laboratories.23
• Wear protective clothing and disposable gloves while handling all reagents.• Wash hands thoroughly after performing the test.• Do not pipette by mouth.• Do not smoke, drink, chew or eat in areas where specimens or kit reagents are being handled.• Dispose of unused reagents and waste in accordance with local, state, provincial and/or federal regulations.• Consult the BD MAX System User’s Manual24 for additional warnings, precautions and procedures.
4
STORAGE AND STABILITY Specimen StabilityCollected specimens should be transferred to the BD MAX UVE Sample Buffer Tubes as prescribed in the Specimen Collection kit package insert. Specimens in BD MAX UVE Sample Buffer Tubes can be stored for a maximum of 8 days at 2–30 °C or for a maximum of 14 days at 2–8 °C before testing. Kit Components StorageBD MAX Vaginal Panel components are stable at 2–25 °C through the stated expiration date. Do not use expired components. BD MAX Vaginal Panel Master Mixes and Extraction Tubes are provided in sealed pouches. To protect product from humidity, immediately re-seal pouches after opening. Reagent tubes are stable for up to 14 days at 2–25 °C after initial opening and re-sealing of the pouch.Unreconstituted BD MAX Master Mixes and Extraction Tubes are stable for up to 24 hours at 2–25 °C after being removed from their protective pouch.
INSTRUCTIONS FOR USESpecimen Transport/PreparationNOTE: Wear gloves when handling the BD MAX UVE Specimen Collection kit components and specimens. If gloves come in contact with the specimen, immediately change them to prevent contamination of other specimens.Specimens should be collected using the BD MAX UVE Specimen Collection kit and following the BD MAX UVE Specimen Collection kit instructions. For specimens not yet transferred into the BD MAX UVE Sample Buffer Tube, prepare the samples for testing following the instructions below. Transfer of Vaginal Swab Specimens to the BD MAX UVE Sample Buffer Tube:NOTE: Swabs must be transferred from the swab sheath to the BD MAX UVE Sample Buffer Tube directly (preferred) or within 2 hours of collection when kept at 2–30 °C.NOTE: Refer to Figure 1 for a summary of the swab specimen transfer workflow.1. Uncap the BD MAX UVE Sample Buffer Tube and fully insert the swab into the tube so that the tip is at the bottom.2. Grasping the swab by the cap, carefully break the swab shaft at the score mark. Use caution to avoid splashing or contamination
of the tube contents.3. Tighten the cap securely on the BD MAX UVE Sample Buffer Tube. In the event that the swab shaft is too long to allow closing
the tube securely, request the collection of a new specimen using a new swab.4. Label the BD MAX UVE Sample Buffer Tube with patient information and date/time collected.NOTE: Be careful not to obscure the barcodes on the tube.5. Proceed directly with Specimen Preparation.
5
Figure 1: Transfer of Swab Specimens to the BD MAX UVE Sample Buffer Tube
1. Fully insert the swab into the tubeso that the tip is at the bottom.
2. Carefully break the shaft at thescore mark.
3. Tightly recap the tube. 4. Label tube with patient information.
Specimen PreparationNOTE: One (1) Septum Cap is required for each specimen and each External Control to be tested.1. Place BD MAX UVE Sample Buffer Tubes in a NALGENE Cryogenic Vial Holder and vortex at maximum speed for 1 minute with
the Multi-Tube Vortexer. If the run cannot be started within 4 hours of this vortexing step, vortex again at maximum speedfor 1 minute with the Multi-Tube Vortexer before starting the run.
2. Uncap the BD MAX UVE Sample Buffer Tube and gently press the swab against the side of the tube to remove excess fluid.NOTE: If a swab is not captured by the cap, do not attempt to remove it from the BD MAX UVE Sample Buffer Tube. The swabmay remain in the BD MAX UVE Sample Buffer Tube for the whole PCR run.
3. Withdraw the swab/cap combination from the tube and discard in a biomedical waste container. Please note that microbialloads from vaginal specimens can be very high, precautions should be taken to avoid environmental contamination during thisparticular step of the Specimen Preparation.
4. Recap the tube with a blue septum cap.5. Proceed directly with BD MAX System Operation. Do not pre-warm tubes prior to using the BD MAX Vaginal Panel.
BD MAX SYSTEM OPERATIONNOTE: Refer to the BD MAX System User’s Manual24 for detailed instructions (Operation section).NOTE: Testing of the BD MAX Vaginal Panel must be performed within 4 hours after the vortexing step above (refer to Specimen Preparation, Steps 1–3). If retesting is necessary, re-vortex sample(s).NOTE: One (1) Vaginitis Master Mix (C4), one (1) Vaginosis Master Mix (C5), one (1) Extraction Tube (C6), and one (1) BD MAX Vaginal Panel Unitized Reagent Strip are required for each specimen and each External Control to be tested.1. Power on the BD MAX System (if not already done) and log in by entering <user name>and <password>.2. Gloves must be changed before manipulating reagents and cartridges.3. Remove the required number of Unitized Reagent Strips from the BD MAX Vaginal Panel kit. Gently tap each Unitized Reagent
Strip onto a hard surface to ensure that all liquids are at the bottom of the tubes.4. Remove the required number of Extraction Tube(s) and Master Mix Tube(s) from their protective pouches. Remove excess air,
and close pouches with the zip seal.
6
5. For each sample to be tested, place 1 Unitized Reagent Strip on the BD MAX System Rack, starting with Position 1 of Rack A.6. Snap 1 Extraction Tube (white foil) into each Unitized Reagent Strip in Position 1 as shown in Figure 2.7. Snap 1 Vaginosis Master Mix Tube (green foil) into each Unitized Reagent Strip in Position 2 as shown in Figure 2.8. Snap 1 Vaginitis Master Mix Tube (blue foil) into each Unitized Reagent Strip in Position 4 as shown in Figure 2.
Figure 2: Snap BD MAX Vaginal Extraction Tubes and Master Mix Tubes into Unitized Reagent Strips.
9. Click on the Run icon, then Inventory. Enter the BD MAX Vaginal Panel kit lot number (for lot traceability) by either scanning the barcode with the scanner or by manual entry.
NOTE: Repeat step 9 each time a new kit lot is used.10. Navigate to the Worklist. Using the pull down menu select <BD MAX Vaginal 46>.11. Enter the Sample Buffer Tube ID, Patient ID and Accession Number (if applicable) into the Worklist, either by scanning the
barcode with the scanner or by manual entry.12. Select the appropriate kit lot number (found on the outer box) from the pull down menu.13. Repeat steps 10 to 12 for all remaining Sample Buffer Tubes.14. Place the Sample Buffer Tubes in the BD MAX System Rack(s) corresponding to the Unitized Reagent Strips assembled in
steps 5 to 8.15. Place the required number of BD MAX PCR Cartridge(s) into the BD MAX System (refer to Figure 3).
• Each cartridge accommodates 12 samples tested against the Vaginosis and the Vaginitis panels for a total of 24 PCR reactions per cartridge.
• The BD MAX System will automatically select the position and row on the BD MAX PCR Cartridge for each run. BD MAX PCR Cartridges may be used multiple times until all lanes have been utilized.
• To maximize use of BD MAX PCR Cartridges, using 2000 Sample Mode, select Run Wizard under the Worklist tab for lane assignments.
• Consult the BD MAX System User’s Manual24 for more details.
Figure 3: Load BD MAX PCR Cartridges
7
16. Load rack(s) into the BD MAX System (refer to Figure 4).
Side A Side B
Figure 4: Load Rack(s) into the BD MAX System.
17. Close the BD MAX System lid and click <Start>to begin processing.18. At the end of the run, check results immediately or store Sample Buffer Tubes according to the temperatures and times stated
in “Repeat Test Procedure” (below) until the results are checked.NOTE: If a septum cap was damaged during the run, replace it with a new one before storing the sample.NOTE: After the end of the run, Sample Buffer Tubes containing samples can be stored:
for a maximum of 5 hours at 2–30 °C or for a maximum of 120 hours at 2–8 °C.
When an Indeterminate (IND), Unresolved (UNR), or Incomplete (INC) result is obtained, or when an External Control failure occurs, a repeat test from the prepared Sample Buffer Tube must be performed (see “Repeat Test Procedure”). If an External Control fails, repeat testing of all specimens using freshly prepared External Controls (see “Quality Control”).
QUALITY CONTROLQuality control procedures monitor the performance of the assay. Laboratories must establish the number, type and frequency of testing control materials according to guidelines or requirements of local, provincial, state, federal and/or country regulations or accreditation organizations. For general Quality Control guidance, the user may wish to refer to Clinical Laboratory Standards Institute documents MM3, C24 and EP12.25,26,27
1. External Control materials are not provided by BD. External Positive and Negative Controls are not used by the BD MAXSystem software for the purpose of sample test result interpretation. External Controls are treated as if they were patientsamples. BD MAX UVE Sample Buffer Tubes are needed to prepare External Controls. (Refer to the table in the ResultsInterpretation section for the interpretation of External Control assay results.)
2. One External Positive Control and one External Negative Control should be run at least daily until adequate process validationis achieved on the BD MAX System in each laboratory setting. Each target control strain should be tested alternately. Reducedfrequency of control testing should be in accordance with applicable regulations.
3. The External Positive Control is intended to monitor for substantial reagent failure. The External Negative Control is intended todetect reagent or environmental contamination (or carry-over) by target nucleic acids.
4. Control strains should be tested according to guidelines or requirements of local, state and/or federal regulations oraccreditation organizations in order to monitor the effectiveness of the entire analytical process.
5. Various types of External Controls are recommended to allow the user to select the most appropriate for their laboratory qualitycontrol program.
a. External Negative Control: A suspension of commercially available control material [e.g., Lactobacillus iners(ATCC® 55195)] in a BD MAX UVE Sample Buffer Tube or a previously characterized sample known to be negative.BD recommends that the External Negative Control be prepared prior to the External Positive Control in order to reducethe potential for contamination as a result of control preparation.
b. External Positive Control: A suspension of the commercially available control materials listed below in a BD MAX UVESample Buffer Tube (Table 1) or a previously characterized sample known to be positive.
8
Table 1: Commercially Available Materials for External Controls
Vaginosis panel BV Positive External Controlaa A mixture of Gardnerella vaginalis and Atopobium vaginae
If control organisms cultures are used:(1) Prepare starting cultures of control organisms as described in the External Control preparation table below.(2) For all controls other than Trichomonas vaginalis, prepare 0.5 McFarland cell suspensions in phosphate buffered saline
(PBS) from fresh culture.(3) Perform serial dilutions listed in Table 2 using PBS to obtain the final dilution recommended for each organism.(4) For BV Positive Control, mix an equal volume of the 0.5 McFarland of Gardnerella vaginalis ATCC 14018 and of the
0.5 McFarland of Atopobium vaginae ATCC BAA-55. Vortex the mix 5–10 sec at maximum speed. No additional mixingis necessary for other controls.
(6) Place BD MAX UVE Sample Buffer Tubes in a NALGENE Cryogenic Vial Holder and vortex at maximum speed for1 minute with the Multi-Tube Vortexer. If the run cannot be started within 4 hours of this vortexing step, vortexagain at maximum speed for 1 minute with the Multi-Tube Vortexer before starting the run.
(7) Process the External Control as if it is a patient sample according to the procedure indicated in the BD MAX SystemOperation section.
Table 2: External Controls Preparation
Controls organism Starting Culture Suspension preparation in PBS Final dilution
Trichomonas vaginalis ATCC 30001 Use ATCC stock From ATCC stock 1/1,000a
Candida albicans ATCC 10231 Fresh culture on Sabouraud plate 0.5 McF in PBS 1/20b
Candida glabrata ATCC 2001 Fresh culture on Sabouraud plate 0.5 McF in PBS 1/100c
Candida krusei ATCC 6258 Fresh culture on Sabouraud plate 0.5 McF in PBS 1/20c
Lactobacillus iners ATCC 55195 Fresh culture on anaerobic blood agar plate 0.5 McF in PBS No dilutionc
Gardnerella vaginalis ATCC 14018 Fresh culture on anaerobic blood agar plate 0.5 McF in PBS No dilution: combine an equal volume of each 0.5 McF
suspensioncAtopobium vaginae ATCC BAA-55 Fresh culture on anaerobic blood agar plate 0.5 McF in PBSa An entire vial of Trichomonas vaginalisATCC30001canbepreparedtothefinaldilution,dividedinto200μLaliquots,frozenandusedforroutinetesting.b Fresh preparation must always be used for Candida albicans ATCC 10231.c Suspensioncanbepreparedtothefinaldilution,dividedinto200μLaliquots,frozenandusedforroutinetesting.
6. External Controls should yield the expected results.a. An External Negative Control that yields a positive test result is indicative of a specimen handling and/or contamination
event. Review the specimen handling technique to avoid mix-up and/or contamination.b. An External Positive Control that yields a negative result is indicative of a specimen handling/ preparation problem.
Review the specimen handling/preparation technique.c. An External Control that yields an Unresolved, Indeterminate or Incomplete test result is indicative of a reagent or a
BD MAX System failure. Check the BD MAX System monitor for any error messages. Refer to the Troubleshootingsection of the BD MAX System User’s Manual24 for interpretation of warning and error codes. If the problem persists,use reagents from an unopened pouch or use a new assay kit.
7. Each Extraction Tube contains a Sample Processing Control which is a plasmid containing a synthetic target DNA sequence.The Sample Processing Control monitors the efficiency of DNA capture, washing and elution during the sample processingsteps, as well as the efficiency of DNA amplification and detection during PCR analysis. If the Sample Processing Controlresult fails to meet the acceptance criteria, the result of the specimen will be reported as Unresolved for this Master Mix.An Unresolved result is indicative of specimen-associated inhibition or reagent failure. Repeat any specimen reported asUnresolved according to the Repeat Test Procedure section below.
9
RESULTS INTERPRETATIONResults are available on the Results tab in the Results window on the BD MAX System monitor. The BD MAX System software automatically interprets test results. A test result may be called NEG (negative), POS (positive) or UNR (unresolved) based on the amplification status of the target and of the Sample Processing Control. IND (indeterminate) or INC (incomplete) results are due to BD MAX System failure. Results interpretation is detailed in Table 3.
Table 3: BD MAX Vaginal Panel Result Interpretation
Assay Result Reported(Vaginitis Master Mix) Interpretation of Results
UNR Unresolved – inhibitory sample or reagent failure; No target detected and no amplification of Sample Processing Control
IND Indeterminate result due to BD MAX System failure (with Warning or Error Codesa)
INC Incomplete Run (with Warning or Error Codesa)
Assay Result Reported(Vaginosis Master Mix) Interpretation of Results
BV POS
Vaginosis Panel DNA detected. Detection of marker combinations related to bacterial vaginosis: Gardnerella vaginalis and/or
L. crispatus and/or L. jensenii and/orAtopobium vaginae and/or
BVAB-2 and/orMegasphaera-1
BV NEG Detection of marker combinations related to normal vaginal flora
UNR Unresolved – inhibitory sample or reagent failure; No target detected and no amplification of Sample Processing Control
IND Indeterminate result due to BD MAX System failure (with Warning or Error Codesa)
INC Incomplete Run (with Warning or Error Codesa)a Refer to the Troubleshooting section of the BD MAX System User’s Manual24 for interpretation of warning and error codes.
REPEAT TEST PROCEDURENOTE: Sufficient volume is available for one repeat test from the Sample Buffer Tube. For Sample Buffer Tubes stored at 2–30 °C, retesting must be initiated within 5 hours of the end of the run. Alternatively, for Sample Buffer Tubes stored at 2–8 °C, retesting may be initiated within 120 hours (5 days) of the end of the run. NOTE: New samples may be tested in the same run with repeat samples.NOTE: In the occurrence of UNR or IND results impacting only one Master Mix, the test must be repeated with both Master Mixes clipped on the strip. In the event that the repeat yields a different reportable result for a given target, any positive result should be reported.Unresolved ResultUnresolved results may be obtained in the event that specimen-associated inhibition or a reagent failure prevents proper target or Sample Processing Control amplification. Sample(s) can be repeated from their corresponding Sample Buffer Tube(s) within the timeframes defined above. Vortex the sample(s) for 1 minute and restart following the BD MAX System Operation section.Indeterminate ResultIndeterminate results may be obtained in the event that a System alert or a reagent failure occurs. Sample(s) can be repeated from their corresponding Sample Buffer Tube(s) within the timeframes defined above. Vortex the sample(s) for 1 minute and restart following the BD MAX System Operation section. For the interpretation of warning or error code messages, refer to the BD MAX System User’s Manual24 (Troubleshooting section).
10
Incomplete ResultIncomplete results may be obtained in the event that Sample Preparation or the PCR failed to complete. Sample(s) can be repeated from their corresponding Sample Buffer Tube(s) within the timeframes defined above. Vortex the sample(s) for 1 minute and restart following BD MAX System Operation section. For the interpretation of warning or error code messages, refer to the BD MAX System User’s Manual24 (Troubleshooting section).External Control FailureExternal Controls should yield expected results when tested. If samples have to be repeated due to an incorrect External Control result, they should be repeated from their Sample Buffer Tubes along with freshly prepared External Controls within the timeframes defined above. Vortex the samples for 1 minute and restart following the BD MAX System Operation section.
LIMITATIONS OF THE PROCEDURE• The BD MAX Vaginal Panel is intended for use only with the BD MAX UVE Specimen Collection kit. • The BD MAX Vaginal Panel should only be used with the BD MAX System by trained laboratory personnel.• The BD MAX Vaginal Panel has not been validated for vaginal swab specimens collected by patients at home.• Collection and testing of patient-collected vaginal swab specimens with the BD MAX Vaginal Panel is not intended to replace
exam by a clinician. Vaginal infections may result from other causes or concurrent infections may occur. • Public health recommendations should be consulted regarding testing for additional sexually transmitted diseases for patients
with a positive result for Bacterial Vaginosis or T. vaginalis with the BD MAX Vaginal Panel. • Additional microorganisms not detected by the BD MAX Vaginal Panel such as Prevotella, Lachnospira, Sneathia, Mobiluncus,
Mycoplasma hominis, and Ureaplasma spp. have also been found in women with bacterial vaginosis, but are less associated with BV due to their relatively low prevalence, sensitivity and/or specificity.3,6,7
• Patients under 18 years old were not evaluated.• A Cgroup positive result can be due to one or multiple Candida species.• Reliable assay results are dependent on adequate specimen collection. Follow the procedures in this package insert and
the BD MAX UVE Specimen Collection Kit. Failure to follow specimen collection instructions may cause an increase in non-reportable results.
• Lubricants or other products containing substances such as carbomers, can increase the non-reportable rate obtained with BD MAX Vaginal Panel.
• Interference with the BD MAX Vaginal Panel was observed in the presence of the following substances: Conceptrol Vaginal Contraceptive Gel, Clotrimazole Vaginal Cream, Monistat 3 Cream, Vagisil Cream, Replens Vaginal Moisturizing Gel, Surgilube, McKesson Lubricating Jelly, Aquasonic Clear, Metronidazole, Leukocytes. The following substances were observed to interfere at levels above the stated concentrations: Preparation H Hemorrhoidal Cream (>0.8 µL/mL), Zovirax Acyclovir 5% Cream (>3.1 µL/mL), VCF Contraceptive Foam (>3.1 µL/mL), KY Jelly Personal Lubricant (>12.5 µL/mL), MUKO Lubricating Jelly (>3.3µL/mL), Whole Blood (>12.5 µL/mL or >1.25% v/v).
• Interference with the BD MAX Vaginal Panel was observed in the presence of the following microorganisms: Lactobacillus amylovorus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus kefirgranum, and Lactobacillus helveticus, that may all be used in probiotics. Interference with Lactobacillus kefiranofaciens has not been determined.
• Cross-reactivity with the BD MAX Vaginal Panel may be observed with the following organisms: Lactobacillus acidophilus, detected in human stool; Trichomonas tenax, a commensal of the oral cavity.
• The following organisms were observed to cross-react above the stated concentrations: Olsenella uli (>6.6 x 104 CFU/mL) and
Atopobium rimae (>4.4 x 104 CFU/mL), both isolated from oral cavity; Lactobacillus delbrueckii subsp. lactis (>3.9 x 103 CFU/mL), detected in human stool; Pichia fermentans (>6.0 x 103 CFU/mL), from coffee beans and fruit.
• All strains tested in the Inclusivity study were detected. However, Three (3) out of 10 tested Gardnerella vaginalis strains and 1 out of 5 Lactobacillus crispatus strains were detected only at high concentrations (9x LoD and 5x LoD respectively).
• The effects of other potential variables such as vaginal discharge, use of tampons and specimen collection variables have not been determined.
• As with many diagnostic tests, results from the BD MAX Vaginal Panel should be interpreted in conjunction with other laboratory and clinical data available to the physician.
• Candida species can be present as commensal organisms in a significant percentage of women and BD MAX Vaginal Panel results should be considered in conjunction with other clinical and patient information to determine the disease status.
• Bacterial vaginosis marker combinations that generate positive results for bacterial vaginosis can be commensal in a significant percentage of women and therefore positive results for bacterial vaginosis should be considered in conjunction with other clinical and patient information to determine the disease status.
• Erroneous test results may occur from improper specimen collection, handling or storage, technical error, sample mix-up or because the number of organisms in the sample is below the analytical sensitivity of the test.
• If the BD MAX Vaginal Panel result is IND, INC, or UNR (for one or more targets) then the test should be repeated.• Good laboratory technique is essential for the proper performance of this assay. Due to the high analytical sensitivity of this test,
extreme care should be taken to preserve the purity of all materials and reagents.• A positive test result does not necessarily indicate the presence of viable organisms. A positive result is indicative of the
presence of target DNA. • The BD MAX Vaginal Panel cannot be used to assess therapeutic success or failure since target nucleic acids may persist
following antimicrobial therapy.
11
• As with all PCR-based tests, extremely low levels of target below the LoD of the assay may be detected, but results may notbe reproducible.
• False negative results may occur due to loss of nucleic acid from inadequate collection, transport or storage of specimens, ordue to inadequate cell lysis. The Sample Processing Control has been added to the test to aid in the identification of specimensthat contain inhibitors to PCR amplification. The Sample Processing Control does not indicate if nucleic acid has been lost dueto inadequate collection, transport or storage of specimens, or if cells have been adequately lysed.
• At very low load, false negative Candida glabrata results may occur for specimens containing more than one analyte.• At very low load, false negative Candida spp. results may occur for specimens containing more than one BV analyte at high loads.• At very low load, false negative Candida spp., C. krusei or C. glabrata results may occur for specimens containing high load
of T. vaginalis.• Mutations or polymorphisms in primer- or probe-binding regions may affect detection resulting in a false negative result with the
BD MAX Vaginal Panel.• This test is a qualitative test and does not provide quantitative values nor indicate the quantity of organisms present.
EXPECTED VALUESIn the BD MAX Vaginal Panel clinical trial, reportable results from specimens compliant at the specimen and PCR levels were obtained from 10 geographically diverse sites. Clinician-collected specimens totaled 1,706 for bacterial vaginosis and 1,704 for other assay targets. Self-collected specimens totaled 1,715 for bacterial vaginosis and 1,714 for other assay targets. The number and percentage of positive cases per target, as determined by the BD MAX Vaginal Panel, are presented in Table 4.
Table 4: BD MAX Vaginal Panel Positivity Rate per Clinic Type
Collection Type Clinic Type Bacterial Vaginosis Cgroupa Candida
glabrataCandida krusei
Trichomonas vaginalis
Clinician-collected
STD / HIV 72.5% (224/309)
34.0% (105/309)
2.6% (8/309)
0.3% (1/309)
15.9% (49/309)
Family Planning 59.7%(682/1,143)
33.1% (378/1,141)
1.1% (13/1,141)
0.1% (1/1,141)
7.4% (85/1,141)
OB/Gyn 20.5% (52/254)
29.5% (75/254)
2.0% (5/254)
0.8% (2/254)
0.4% (1/254)
Total 56.2%(958/1,706)
32.7% (558/1,704)
1.5% (26/1,704)
0.2% (4/1,704)
7.9% (135/1,704)
Self-collected
STD / HIV 74.4%(229/308)
33.6% (103/307)
2.3% (7/307)
0.0% (0/307)
16.0% (49/307)
Family Planning 59.7%(687/1,151)
35.0% (403/1,151)
1.7% (19/1,151)
0.0% (0/1,151)
7.6% (88/1,151)
OB/Gyn 21.9% (56/256)
34.4% (88/256)
2.3% (6/256)
0.0% (0/256)
0.4% (1/256)
Total 56.7% (972/1,715)
34.7% (594/1,714)
1.9% (32/1,714)
0.0% (0/1,714)
8.1% (138/1,714)
a Candida albicans, Candida tropicalis, Candida parapsilosis and/or Candida dubliniensis
PERFORMANCE CHARACTERISTICS Assay performance characteristics were evaluated in a study performed at 10 geographically diverse specimen collection sites, including 7 performing collection only and 3 also performing BD MAX Vaginal Panel testing. Two (2) reference laboratories performed BD MAX Vaginal Panel and/or reference method testing. For consented adult female subjects presenting with symptoms of vaginitis or bacterial vaginosis, one self-collected and one clinician-collected vaginal swab were collected using the BD MAX UVE Specimen Collection Kit and tested independently. Three (3) additional vaginal swabs were collected for reference method testing. Three (3) reference methods were performed for each patient:1) Bacterial vaginosis status was determined using a combination of Nugent Score and Amsel’s criteria. Specimens with normal flora
as per the Nugent Score were considered negative; those with BV flora were considered positive while those with intermediateflora were segregated in positive or negative categories using Amsel’s criteria. Samples positive for 2 out of the 3 following criteriawere considered Amsel’s positive: vaginal pH >4.5, presence of clue cells and positive Whiff test.
2) Candida status was determined by selective (Candida) chromogenic medium and Sabouraud Dextrose Emmons plate culturefollowed by PCR amplification and bi-directional sequencing of the its2 gene for Candida species identification.
3) Trichomonas vaginalis status was determined by microscopic visualization of motile trichomonads in saline wet mount ofvaginal secretion and by culture. Any positive result was sufficient to consider the specimen reference method positive forTrichomonas vaginalis.
A total of 1,763 subjects were enrolled in the study. Of those, 1,740 subjects were compliant and 23 were found non-compliant as per protocol criteria. For clinician-collected specimens, the numbers of compliant specimens with reportable reference method and BD MAX Vaginal Panel results were 1,579 for bacterial vaginosis, 1,621 for Candida and 1,604 for Trichomonas vaginalis. For self-collected specimens, the numbers of compliant specimens with reportable reference method and BD MAX Vaginal Panel results were 1,589 for bacterial vaginosis, 1,632 for Candida and 1,615 for Trichomonas vaginalis.
12
BV Performance Results Bacterial vaginosis per site and overall performance results are presented in Table 5. The sensitivity and specificity were 90.1 and 86.2% respectively for clinician-collected vaginal swabs, and 90.5 and 84.6% respectively for self-collected vaginal swabs. For the population tested, this resulted in Positive Predictive Values (PPV) of 89.2 and 88.2% for clinician- and self-collected specimens, respectively. Negative Predictive Values (NPV) of 87.4% and 87.6% were obtained for clinician- and self-collected specimens, respectively. BV prevalence was 55.8% for patients with compliant reference method results.
Table 5: BV Performance Results per Collection Type and Collection Site
Table 6 shows BV performance stratified by age groups. All age groups showed sensitivities between 89.5% and 94.9% except for patients aged 50 and over, which displayed sensitivity of 68.8% and 62.5% for clinician-collected and self-collected specimens, respectively. Specificity varies from 84.7% to 91.4% for clinician-collected specimens and 83.1% to 91.7% for self-collected specimens.
Table 7 shows BV performance stratified by ethnicity. For clinician-collected specimens, sensitivity varies from 79.3% to 91.4% and specificity varies from 79.7% to 100% across the most prevalent ethnic groups in the clinical trial. PPV varies from 78.3% to 100% and NPV varies from 82.4% to 93.4%. For self-collected specimens, sensitivity varies from 79.3% to 92.2% and specificity varies from 77.1% to 90.5% across the most prevalent ethnic groups. PPV varies from 81.4% to 88.3% and NPV varies from 80.6% to 92.9%.
Table 7: BV Performance Results Stratified per Ethnicity
(80.5, 93.8) (80.5, 95.5) (86.5, 97.0) (76.8, 91.8) (80.5, 93.8) (82.5, 96.5) (87.8, 97.8) (77.0, 91.8)a Prevalence was calculated for specimens with compliant reference method results. b CI: Confidence interval
14
Table 8 shows BV performance stratified by subgroups relevant to health condition. For clinician-collected specimens, sensitivity varies from 78.9% to 90.5% and specificity varies from 68.3% to 94.0% across subgroups. For self-collected specimens, sensitivity varies from 78.6% to 91.0% and specificity varies from 69.8% to 90.9% across subgroups.
Table 8. BV Performance Stratified by Subgroups Relevant to Health Condition
Patients with unprotected intercourse in the last 24 h
89.7 61/68
(80.2, 94.9)
68.3 28/41
(53.0, 80.4)
88.260/68
(78.5, 93.9)
69.830/43
(54.9, 81.4)
Patients with recurrent symptoms 87.6
162/185(82.0, 91.6)
87.5161/184
(81.9, 91.5)
86.1161/187
(80.4, 90.3)
86.0160/186
(80.3, 90.3)
Patients using oral antibiotics82.3
79/96(73.5, 88.6)
94.078/83
(86.7, 97.4)
84.481/96
(75.8, 90.3)
83.370/84
(73.9, 89.8)
Patients with menses83.3
40/48(70.4, 91.3)
86.833/38
(72.7, 94.2)
85.7 42/49
(73.3, 92.9)
86.8 33/38
(72.7, 94.2)
Patients without menses90.5
753/832(88.3, 92.3)
86.1563/654
(83.2, 88.5)
90.8758/835
(88.6, 92.6)
84.4557/660
(81.4, 87.0)a CI: Confidence interval
15
Cgroup Performance Results Cgroup (Candida albicans, Candida tropicalis, Candida parapsilosis and/or Candida dubliniensis) per site and overall performance results are presented in Table 9. The sensitivity and specificity were 90.9 and 94.2% respectively for clinician-collected vaginal swabs, and 92.0 and 92.1% respectively for self-collected vaginal swabs. For the population tested, this resulted in PPV of 88.0 and 84.2% for clinician- and self-collected specimens, respectively. NPV of 95.8 and 96.1% were obtained for clinician-collected vaginal swabs, respectively. The prevalence of those Candida species combined was 31.6% for patients with compliant reference method results.
Table 9: Cgroup Performance Results per Collection Type and Collection Site
In the BD MAX Vaginal Panel, Cgroup includes Candida albicans, Candida tropicalis, Candida parapsilosis and/or Candida dubliniensis. Table 10 shows the sensitivity of the BD MAX Vaginal Panel for each Cgroup species.
Table 10: Cgroup Performance Results Stratified by Candida Species Detected
Species (its2 gene ID)
SensitivityClinician-collected Self-collected
Estimate95% CI
Candida albicans91.0%
445/489 (88.1%, 93.2%)
91.9%451/491
(89.1%, 94.0%)
Candida albicans (co-infected with C. glabrata)92.3%12/13
(66.7%, 98.6%)
100%13/13
(77.2%, 100%)
Co-infection Candida albicans and Candida tropicalis 100%
1/1 (20.7%, 100%)
100% 1/1
(20.7%, 100%)
Candida dubliniensis100%
3/3 (43.9%, 100%)
100% 3/3
(43.9%, 100%)
Candida tropicalis50.0%
1/2 (9.5%, 90.5%)
66.7% 2/3
(20.8%, 93.9%)
Overall90.9
462/508(88.1, 93.1)
92.0470/511
(89.3, 94.0)
Table 11 shows Cgroup performance stratified by age groups. Sensitivity varies from 77.8% to 93.9% for clinician-collected specimens and 88.4% to 100% for self-collected specimens. Specificity varies from 93.7% to 100% for clinician-collected specimens and 91.5% to 98.3% for self-collected specimens.
Table 11: Cgroup Performance Results per Age Group
Table 12 shows Cgroup performance stratified by ethnicity. For clinician-collected specimens, sensitivity varies from 88.8% to 92.2% and specificity varies from 93.0% to 97.8% across the most prevalent ethnicities in the clinical trial. PPV varies from 86.3% to 94.0% and NPV varies from 95.0% to 97.7%. For self-collected specimens, sensitivity varies from 90.9% to 92.4% and specificity varies from 90.4% to 93.8% across the most prevalent subgroups. PPV varies from 78.4% to 87.2% and NPV varies from 95.5% to 97.6%.
Table 12: Cgroup Performance Results per Ethnicity
(81.8, 97.7) (89.9, 98.1) (79.1, 95.9) (92.9, 99.4) (78.8, 96.4) (86.5, 96.3) (73.2, 91.7) (91.7, 98.9)a Prevalence was calculated for specimens with compliant reference method results.b CI: Confidence interval
Table 13 shows Cgroup performance stratified by subgroups relevant to health condition. For clinician-collected specimens, sensitivity varies from 85.7% to 100% and specificity varies from 88.9% to 94.3% across subgroups. For self-collected specimens, sensitivity varies from 90.0% to 98.1% and specificity varies from 86.0% to 95.5% across subgroups.
Table 13. Cgroup Performance Stratified by Subgroups Relevant to Health Condition
Patients with unprotected intercourse in the last 24 h
94.7 36/38
(82.7, 98.5)
93.1 67/72
(84.8, 97.0)
95.0 38/40
(83.5, 98.6)
93.1 67/72
(84.8, 97.0)
Patients with recurrent symptoms 90.3
93/103(83.0, 94.6)
92.7 253/273
(89.0, 95.2)
93.5 100/107
(87.1, 96.8)
91.2249/273
(87.3, 94.0)
Patients using oral antibiotics94.9
56/59(86.1, 98.3)
92.2 119/129
(86.3, 95.7)
96.6 57/59
(88.5, 99.1)
86.9113/130
(80.1, 91.7)
Patients with menses85.7
18/21(65.4, 95.0)
94.0 63/67
(85.6, 97.7)
95.5 21/22
(78.2, 99.2)
95.5 64/67
(87.6, 98.5)
Patients without menses91.3
443/485(88.5, 93.5)
94.3982/1,041
(92.8, 95.6)
92.0448/487
(89.2, 94.1)
91.8963/1,049
(90.0, 93.3)a CI: Confidence interval
18
Candida glabrata Performance Results Candida glabrata per site and overall performance results are presented in Table 14. The sensitivity and specificity were 75.9 and 99.7% respectively for clinician-collected vaginal swabs and 86.7 and 99.6% respectively for self-collected vaginal swabs. For the population tested, this resulted in PPV of 84.8 and 81.0% for clinician- and self-collected specimens, respectively. NPV of 99.6 and 99.8% were obtained for clinician- and self-collected specimens, respectively. The prevalence of C. glabrata was 1.8% for patients with compliant reference method results.As Candida glabrata’s prevalence is low, an evaluation of contrived specimens was performed to supplement data collected in the study. These were prepared by spiking 50 different Candida glabrata strains in individual vaginal matrices. True negative specimens, containing vaginal matrix only, were also tested. Strains were spiked at various clinically relevant loads and randomly distributed among 3 study testing sites for BD MAX Vaginal Panel testing. A positive agreement of 100% was obtained across the tested loads. Results are shown in Table 15.
Table 14: Candida glabrata Overall Performance Results per Collection Type and Collection Site
a CI: Confidence intervalb Out of 7 C. glabrata false negative results, 6 showed chromagar results consistent with low C. glabrata load (1+ to 2+ growth level) and 1 showed chromagar
result consistent with high C. glabrata load (3+ growth level)c The BD MAX Vaginal Panel detected BV and/or Cgroup signals in 6 out of 7 specimens with C. glabrata false negative resultsd Out of 4 C. glabrata false negative results, 3 showed chromagar results consistent with low C. glabrata load (1+ to 2+ growth level) and 1 showed chromagar
result consistent with high C. glabrata load (3+ growth level)e The BD MAX Vaginal Panel detected BV and/or Cgroup signals in the 4 specimens with C. glabrata false negative results
19
Table 15: Candida glabrata Contrived Specimens Results per Category
Candida glabrata Percent Agreement
Category Load (x LoD) Percent(95% CIa)
High Positive ≥10and<20100(5/5)
(56.6, 100)
Moderate Positive ≥2and<10100
(20/20)(83.9, 100)
Low Positive ≥1and<2100
(25/25)(86.7, 100)
True Negative 0100
(50/50)(92.9, 100)
a CI: Confidence interval
Table 16 shows Candida glabrata performance stratified by age groups. Sensitivity varies from 66.7% to 100% for clinician-collected specimens and 78.9% to 100% for self-collected specimens. Specificity varies from 99.4% to 100% across age groups and collection types.
Table 16: Candida glabrata Performance Results per Age Group
Table 17 shows Candida glabrata performance stratified by ethnicity. For clinician-collected and self-collected specimens, sensitivity varies from 50.0% to 100% and specificity varies from 99.3% to 100% across ethnicities.
Table 17: Candida glabrata Performance Results per Ethnicity
(51.0, 100) (96.3, 99.9) (39.7, 99.4) (98.4, 100) (51.0, 100) (96.4, 99.9) (39.9, 99.4) (98.4, 100)a Prevalence was calculated for specimens with compliant reference method results.b CI: Confidence interval
Table 18 shows Candida glabrata performance in pregnant patients. There was no data for sensitivity calculation and specificity is 100% for both clinician- and self-collected specimens.
Table 18. Candida glabrata Performance in Pregnant Patients
Pregnant patients No data for Sensitivity calculation
100 20/20
(83.9, 100)
No data for Sensitivity calculation
100 19/19
(83.2, 100)a CI: Confidence interval
21
Candida krusei Performance Results Candida krusei overall performance results are presented in Table 19. No Candida krusei positive specimens were identified in the study by the reference method; thus no data is available for sensitivity calculation. The specificity was 99.8 and 100% for clinician-collected and self-collected vaginal swabs, respectively. The PPV and NPV cannot be calculated since the prevalence value was zero.As Candida krusei is a rare analyte, an evaluation of contrived specimens was performed to supplement data collected in the study. These were prepared by spiking 50 different Candida krusei strains in individual vaginal matrices. True negative specimens, containing vaginal matrix only, were also tested. Strains were spiked at various clinically relevant loads and randomly distributed among 3 study testing sites for BD MAX Vaginal Panel testing. A positive agreement of 100% was obtained across the tested loads. Results are shown in Table 20.
Table 19: Candida krusei Overall Performance Results per Collection Type
Collection TypeSensitivity Specificity
Percent(95% CIa)
Percent(95% CIa)
Clinician-collected No data for Sensitivity calculation99.8
1,617/1,621(99.4, 99.9)
Self-collected No data for Sensitivity calculation100
1,632/1,632(99.8, 100)
a CI: Confidence interval
Table 20: Candida krusei Contrived Specimens Results per Category
Candida krusei Percent Agreement
Category Load (x LoD) Percent(95% CIa)
High Positive ≥10and<201005/5
(56.6, 100)
Moderate Positive ≥2and<10100
20/20(83.9, 100)
Low Positive ≥1and<2100
25/25(86.7, 100)
True Negative 0100
50/50(92.9, 100)
a CI: Confidence interval
22
Trichomonas vaginalis Performance Results Trichomonas vaginalis per site and overall performance results are presented in Table 21. The sensitivity and specificity were 93.1 and 99.3% respectively for clinician-collected vaginal swabs and 93.2 and 99.3% respectively for self-collected vaginal swabs. For the population tested, this resulted in PPV of 91.8% and 91.9% for clinician- and self-collected specimens, respectively, and NPV of 99.4 for both collection types. The prevalence of T. vaginalis was 8.2% for patients with compliant reference method results.
Table 21: Trichomonas vaginalis Performance Results per Collection Type and Collection Site
(98.7, 99.6)a CI: Confidence intervalb 9 false-negative results were recorded. Of those, 7 were found negative with an FDA-cleared molecular method.c 11 false-positive results were recorded. Of those, 10 were found positive with an FDA-cleared molecular method.
23
Table 22 shows Trichomonas vaginalis performance stratified by age groups. All age groups show sensitivities of 93.8% or above except for patients aged 50 and over, which display sensitivity of 66.7% for both collection types. Specificity of 98.2% or above was obtained across age groups and collection types.
Table 22: Trichomonas vaginalis Performance Results per Age Group
Table 23 shows Trichomonas vaginalis performance stratified by ethnicity. For clinician-collected specimens, sensitivity varies from 70.0% to 100% and specificity varies from 98.2% to 99.5% across ethnicities. For self-collected specimens, sensitivity varies from 70.0% to 100% and specificity varies from 98.4% to 100% across ethnicities.
Table 23: Trichomonas vaginalis Performance Results per Ethnicity
(95.1, 99.4)a Prevalence was calculated for specimens with compliant reference method results.b CI: Confidence interval
24
Table 24 shows Trichomonas vaginalis performance stratified by subgroups related to health condition. For clinician-collected specimens, sensitivity varies from 92.0% to 100% and specificity varies from 99.7% to 100% across subgroups. For self-collected specimens, sensitivity varies from 92.3% to 100% and specificity varies from 99.4% to 100% across subgroups.
Table 24. Trichomonas vaginalis Performance Stratified by Subgroups Related to Health Condition
Co-Infection RateCo-infections calculation based on specimens that were compliant for all targets and for both BD MAX Vaginal Panel and Reference Method are reported in Table 25. The most prevalent co-infection was a combination of BV and Cgroup with 13.8% and 15.2% for clinician- and self-collected specimens respectively. In total, 21.2% and 23.1% of clinician-collected and self-collected specimens, respectively, showed co-infection.
Table 25. BD MAX Vaginal Panel Co-detection Rates
Co-Infection Clinician-collected Self-collected
BV and Cgroup 13.8%206/1,493
15.2%228/1,504
BV and TV 4.8%72/1,493
4.5%68/1,504
BV and Cgroup and TV 1.4%21/1,493
1.5%23/1,504
BV and Cgroup and Cgla 0.4%6/1,493
0.6%9/1,504
BV and Cgla 0.2%3/1,493
0.5%7/1,504
Cgroup and TV 0.3%4/1,493
0.3%5/1,504
Cgroup and Cgla 0.2%3/1,493
0.4%6/1,504
BV and Cgroup and Ckru 0.1%2/1,493
0.0%0/1,504
BV and Cgla and TV 0.0%0/1,493
0.1%1/1,504
Total 21.2%317/1,493
23.1%347/1,504
25
A comparison of co-infections based on all reportable specimen results is presented in Table 26. Numbers are presented for clinician/self-collected specimens. Bolded numbers represent co-infection events with concordant reference method and BD MAX Vaginal Panel results. Non-bolded entries represent specimens with discordant results. Concordant single infections are not represented.
Table 26. Co-Infections Observed during the BD MAX Vaginal Panel Clinical Trial
Total Number of Occurrences Clinician collected / Self- collectedComposite Reference Method
Non-Reportable RateOf all the specimens initially evaluated with the BD MAX Vaginal Panel, 0.3 and 0.7% initially reported as Unresolved for clinician- and self-collected specimens, respectively. Following a valid repeat test, 0.2 and 0.1% remained Unresolved for clinician- and self-collected specimens, respectively. Of all the specimens initially evaluated with the BD MAX Vaginal Panel, 3.7 and 2.7% initially reported as Indeterminate for clinician- and self-collected specimens, respectively. Following a valid repeat test, 0.7 and 0.5% remained Indeterminate for clinician- and self-collected specimens, respectively. Of all the specimens initially evaluated with the BD MAX Vaginal Panel, 1.4% initially reported as Incomplete for both collection types. Following a valid repeat test, 0.2% remained Incomplete for both collection types. The total rates of non-reportable results were 5.4 and 4.8% for clinician- and self-collected specimens, respectively. Following a valid repeat test, 1.0 and 0.8% remained non-reportable for clinician- and self-collected specimens, respectively. Results are shown in Table 27.
Table 27: Non-reportable Rates
Collection Type
Unresolved Rate Indeterminate Rate Incomplete Rate Total RateInitial
Percent(95% CIb)
FinalaPercent
(95% CIb)
InitialPercent
(95% CIb)
FinalaPercent
(95% CIb)
InitialPercent
(95% CIb)
FinalaPercent
(95% CIb)
InitialPercent
(95% CIb)
FinalaPercent
(95% CIb)
Clinician-collected
0.3 6/1,734
(0.2, 0.8)
0.23/1,729
(0.1, 0.5)
3.764/1,734(2.9, 4.7)
0.712/1,729(0.4, 1.2)
1.424/1,734(0.9, 2.1)
0.23/1,729
(0.1, 0.5)
5.494/1,734(4.5, 6.6)
1.018/1,729(0.7, 1.6)
Self-collected0.7
12/1,736(0.4, 1.2)
0.11/1,735
(0.0, 0.3)
2.747/1,736(2.0, 3.6)
0.59/1,735
(0.3, 1.0)
1.424/1,736(0.9, 2.0)
0.24/1,735
(0.1, 0.6)
4.883/1,736(3.9, 5.9)
0.814/1,735 (0.5,1.3)
a The final rate is calculated with valid repeats onlyb CI: Confidence interval
26
Positivity Rates for the BD MAX Vaginal Panel in an Asymptomatic PopulationAlthough the BD MAX Vaginal Panel is not intended for testing specimens from asymptomatic women, presence of Candida species, T. vaginalis, and BV markers has been reported in this population.28,29,30 Presence of Candida species, T. vaginalis, and BV markers was evaluated on 202 asymptomatic women using the BD MAX Vaginal Panel (Table 28). All vaginitis targets were detected by the BD MAX Vaginal Panel, with rates varying from 1.5% for C. krusei to 20.8% for Cgroup. BV signals were also detected in 33.7% of asymptomatic women. Table 28 also displays results for the most prevalent ethnic groups in the study. BV, Cgroup, and T. vaginalis were detected in all ethnic categories.
Table 28. BD MAX Vaginal Panel Positive Rates in Asymptomatic Women
Target OverallBy Ethnic Group
Black/African American White (not Hispanic) Othersa
BV 33.7%68/202
39.4%(37/94)
28.8%(23/80)
28.6%(8/28)
Cgroup20.8%42/202
22.3% (21/94)
16.3% (13/80)
28.6%(8/28)
C. glabrata5.9%
12/20211.7% (11/94)
0.0% (0/80)
3.6%(1/28)
C. krusei1.5%3/202
1.1% (1/94)
2.5% (2/80)
0.0%(0/28)
T. vaginalis11.4%23/202
22.3% (21/94)
1.3% (1/80)
3.6%(1/28)
a Including: American Indian or Alaska natives, Asian, Mixed Ethnicity and Unknown
Analytical sensitivityThe analytical sensitivity (Limit of detection or LoD) for the BD MAX Vaginal Panel was determined as follows: Representative bacterial suspensions were prepared for each of the target organisms detected by the BD MAX Vaginal Panel. Positive specimens were prepared by inoculating simulated vaginal matrix in BD MAX UVE Sample Buffer with multiple concentrations of each representative strain. The LoD for each vaginitis strain was then confirmed in natural matrix. The LoD for each vaginosis strain was confirmed in simulated matrix. Table 29 lists the confirmed LoD values of the Vaginitis Master Mix targets and Vaginosis Master Mix markers.
Analytical InclusivityAn analytical inclusivity study was performed using a variety of BD MAX Vaginal Panel target strains commonly found in vaginal specimens, taking into account phylogenetic diversity, geographic origin and temporal diversity. Five of each Candida strain (C. albicans, C. dubliniensis, C. parapsilosis, C. tropicalis, C. glabrata, and C. krusei) and nine Trichomonas vaginalis strains, as well as five Atopobium vaginae strains, ten strains of Gardnerella vaginalis, five Lactobacillus crispatus strains and five Lactobacillus jensenii strains were tested, including strains from public collections and well-characterized clinical isolates. These 64strainswereinoculatedat<3xLoDofthecorrespondingreferencestrain.TheBDMAXVaginalPanelcorrectlyidentified59ofthe strains tested upon initial testing. Four strains of Gardnerella vaginalis and one strain of Lactobacillus crispatus did not meet acceptance criteria and were further evaluated. One Gardnerella vaginalis strain was correctly identified after repeat at the initial concentration and the other strains were titrated and tested to determine the minimum concentration sufficient for detection. Upon repeat, one G. vaginalisstrainwasdetectedat<3xLoD.Thethreeothersweredetectedat~9xLoD.TheL. crispatus strain was detectedat~5xLoD.Analytical Specificity/Cross-ReactivityThe BD MAX Vaginal Panel was tested with samples containing phylogenetically related species and other organisms likely to be found in vaginal specimens. The bacterial cells, yeasts, parasites and viruses were tested in the BD MAX UVE Sample Buffer Tube at≥106bacteria,cellsorgenomeequivalents/mL,or≥105 PFU/mL or TCID50/mL or equivalent amount of RNA/DNA/PCR reaction. Overall 118 organisms were tested and are listed in Table 30.• Most of bacterial strains, yeast, parasites and viruses tested produced negative results with the BD MAX Vaginal Panel. No
cross-reaction was observed for Gardnerella vaginalis, Megasphaera-1 or BVAB-2.• Candida guillermondii, Candida haemulonii and Candida orthopsilosis, occasionally reported as causes of vulvovaginal
candidiasis,7 produced positive results for Cgroup. No positive result was recorded at Candida guillermondii concentrations ≤6.0x103 CFU/mL.
• Trichomonas tenax (commensal of the oral cavity) produced positive results for Trichomonas vaginalis. • Pichia fermentans (found in coffee beans and fruit) produced positive results for C. krusei. No positive result was recorded at
≤6.0x103 CFU/mL.• Olsenella uli and Atopobium rimae, both isolated from oral cavity, cross-reacted with the bacterial vaginosis marker A. vaginae;
Lactobacillus acidophilus, detected in human stool, cross-reacted with the bacterial vaginosis marker Lactobacillus. No positive result was recorded at Olsenella uliconcentrations≤6.6x104 CFU/mL and Atopobium rimaeconcentrations≤4.4x104 CFU/mL.
• Lactobacillus delbrueckii subsp. Lactis, detected in human stool, cross-reacted with the bacterial vaginosis marker Lactobacillus.Nopositiveresultwasrecordedatconcentrations≤3.9x103 CFU/mL. In addition, at first testing (≥106 bacteria/mL), one Cgroup signal was recorded; the repeat was negative for Cgroup. Ten (10) additional replicates were testedat≥106 bacteria/mL. All cross-reacted with bacterial vaginosis but yielded Cgroup negative results.
• Bifidobacterium breve, E. coli GC10, and Lactobacillus acetotolerans cross-reacted with the bacterial vaginosis marker A. vaginae. Each sample was repeated and yielded a negative result. Ten (10) additional replicates of each strain were tested and all yielded A. vaginae negative results. Among them, one E. coli GC10 replicate yielded a positive signal for Megaspheara-1/BVAB-2. The repeat was all-negative.
• Chlamydia trachomatis yielded a Cgroup signal; the repeat was negative. Ten (10) additional replicates were tested and all yielded negative results.
28
Table 30. Organisms Tested to Determine BD MAX Vaginal Panel Specificity/Cross-Reactivity
Lactobacillus acidophilusa Also reported as Pichia ohmeri, C. guilliermondii b Also reported as C. sorbosac Also reported as C. norvegensisd Candida metapsilosis and Candida orthopsilosis are both subgroups of C. parapsilosis, which is a target of the assay. These two targets were predicted to
cross-react based on in silico analysis. Candida haemulonii, Candida metapsilosis, and Candida orthopsilosis may all be associated with candidiasis.
29
Interfering SubstancesTwenty-four (24) biological and chemical substances that may be present in vaginal specimens were evaluated for potential interference with the BD MAX Vaginal Panel. Exogenous (e.g., prescription and Over-the-Counter drugs, creams and/or gels) and endogenous (e.g., blood, hormones, mucus) were tested at the highest concentrations that may be found in vaginal specimens. KY Jelly Personal Lubricant and Whole Blood were found to interfere at levels above >12.5 µL/mL. Zovirax Acyclovir 5% Cream and VCF Contraceptive Foam were found to interfere at levels above >3.1 µL/mL. MUKO Lubricating Jelly (Gel) was found to interfere above >3.3µL/mL. Preparation H Hemorrhoidal Cream was found to interfere above >0.8 µL/mL. Interference with the following substances was observed at all tested levels: Conceptrol Vaginal Contraceptive Gel, Clotrimazole Vaginal Cream, Monistat 3 Cream, Vagisil Cream, Replens Vaginal Moisturizing Gel, Surgilube (Gel), Aquasonic Clear (Gel), McKesson Lubricating Jelly (Gel), Metronidazole, Leukocytes (Table 31). Substances that demonstrated interference may result in potential unresolved, indeterminate or false negative results.In addition, microorganisms that may be used in probiotics were evaluated for potential interference with the BD MAX Vaginal Panel. Fourteen (14) organisms were tested at high concentrations (>6.7 x 105 CFU/mL of Sample Buffer). Interference in the form of false negative BV results was observed in the presence of the following organisms: Lactobacillus amylovorus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus kefirgranum, and Lactobacillus helveticus. Results are shown in Table 32.
Table 31. Exogenous and Endogenous Substances Tested for Interferenceaa
No Interference Observed Interference Observed
Substance Substance Level Below Which No Interference Observed (µL/mL)
VCF Contraceptive Film Zovirax, Acyclovir 5% Cream ≤3.1
Summer’s Eve Douche Preparation H, Hemorrhoidal Cream ≤0.8
FDS Feminine Deodorant Spray KY Jelly Personal Lubricant ≤12.5
Progesterone MUKO Lubricating Jelly ≤3.3
Estradiol Conceptrol Vaginal Contraceptive Gel
Interference observed at each level evaluated
Clotrimazole Vaginal Cream, USP 2%
Monistat 3 Cream, Miconazole Nitrate, 4%
Vagisil, Benzocaine 20%, Resorcinol 3%
Replens Vaginal Moisturizing Gel
Metronidazole 0.75% Gel
Surgilube
Aquasonic Clear
McKesson Lubricating Jelly
Endo
geno
us Mucus (Bovine Cervical, 5% v/v) Whole Blood ≤12.5(equivalentto≤1.25%v/v)
Semen (5% v/v) Leukocytes Interference observed at each level evaluated
a In total, with the BD MAX Vaginal Panel in the presence of potentially interfering substances, 3,573 samples were tested for vaginosis targets and 4,158 for vaginitis targets. For vaginitis targets, rates of 10.13% IND and 2.86% UNR results were recorded. For BV, rates of 11.67% IND and 0.76% UNR results were recorded.
Table 32. Microorganisms Tested for Interference
No Interference Observed Interference Observed
Lactobacillus plantarum Lactobacillus casei Lactobacillus delbrueckii subsp. bulgaricus
Mixed Infection/Competitive InterferenceA mixed infection/competitive interference study was designed to evaluate the ability of the BD MAX Vaginal Panel to detect low positives in the presence of other targets at high concentrations. The following organisms and concentrations were evaluated.
• Assay targets at high concentrations: L. crispatus (8.7 x 104 CFU/mL), G. vaginalis (5.0 x 106 CFU/mL), A. vaginae(8.0 x 105 CFU/mL), Megasphaera-1 (2.4 x 107 cp/mL) and T. vaginalis (3.3 x 105 to 1.0 x 106 cells/mL), C. albicans(1 x 106 CFU/mL), C. glabrata (1 x 106 CFU/mL).
• High concentrations of organisms of the vaginal flora: Dialister microaerophilus, Prevotella melaninogenica, Streptococcusmitis, Bifidobacterium breve, and Mobiluncus curtisii (each at 1.0 x 106 CFU/mL).
• LowpositiveloadsofVaginitistargetsweretestedat<2xLoD.Samplescontaininglowpositiveloadsofbacterialvaginosismarkers were prepared with organism compositions sufficient to obtain 95% positive BV results.
A series of pools simulating mixed infections were tested: • BV analytes at high loads with low positive loads of:
○ T. vaginalis and C. albicans○ C. krusei and C. glabrata
• C. albicans high load with low positive loads of:○ C. krusei and C. glabrata○ T. vaginalis○ BV
• C. glabrata high load with low positive loads of:○ T. vaginalis○ BV
• C. krusei high load with low positive load of BV• T. vaginalis high load with low positive loads of:
○ BV○ C. albicans○ C. krusei○ C. glabrata
• Vaginal flora high load with low positive loads of:○ T. vaginalis and BV○ C. albicans and BV○ C. glabrata and BV○ C. krusei and BV
Samples containing T. vaginalis at low concentrations and low positive BV samples were successfully detected by the BD MAX Vaginal Panel when combined with high concentrations of other assay targets or with other organisms of the vaginal flora in the presence of simulated vaginal matrix. Competitive inhibition was observed for the following conditions:
• For low positive samples containing Candida albicans, 92% of positive results were obtained in presence of BV analytes athigh loads.
• For low positive samples containing Candida albicans, C. krusei or C. glabrata, BD MAX Vaginal Panel generated 42%,61% and 33% of expected results respectively in presence of Trichomonas vaginalis at a load of 3.3 x 105 cells/mL.
31
PrecisionTo evaluate the BD MAX Vaginal Panel within–laboratory precision, testing was performed at one site with two panels over 12 days per panel. Two operators performed two runs per day, for a total of 48 runs. Panel members were made of target organisms (or plasmid DNA for Megasphaera-1 and BVAB-2) spiked in simulated vaginal matrix. Bacterial vaginosis panel members were prepared using varying concentrations of multiple targeted species with sample compositions designed to generate low positive, moderate positive, high negative, or negative results for bacterial vaginosis. For Candida and Trichomonas vaginalis panel members, the target organisms were spiked at concentrations based on the assay LoD. Table 33 describes the panel members evaluated.
Table 33. Precision Study Spiking Concentrations
Concentration DesignationBacterial Vaginosis Candida and Trichomonas vaginalis
(% of positive results expected based on the organism composition) (x LoD)
Moderate Positive ~100 ≥2to≤5
Low Positive ~95 <2
High BV Negative ~20–80
BV Negative <5
True Negative 0 (No Target) No Target
Precision results are reported in Table 34.
Table 34. Qualitative Precision Study Results Summary- Vaginitis
ConcentrationPercent Agreement with Expected Result
a The expected assay results were deemed to be negative.b Performance includes combined results from replicates of six panel members containing different organism compositions.c Performance includes combined results from replicates of four panel members containing different organism compositions.
ReproducibilityThe reproducibility study was performed using the same sample categories as defined above for the precision study (Table 33) except for the high negative category which was not tested.To evaluate the BD MAX Vaginal Panel site-to-site reproducibility, testing was performed at 3 sites over 8 days. At each site, 2 operators performed 2 runs on alternate days, for a total of 48 runs. Reproducibility results are summarized in Table 35. The qualitative and quantitative reproducibility results across sites and by target are presented in Tables 36–37. Second Derivative Peak Abscissa (SDPA), an internal criterion used to determine a final assay result, was selected as a means of assessing quantitative assay reproducibility. Mean SDPA values with variance components (SD and % CV) are shown in Table 40.
32
Table 35. Qualitative Reproducibility Study Results Summary – Site to Site
ConcentrationPercent Agreement with Expected Result
a The expected assay results were deemed to be negative.b Performance includes combined results from replicates of two panel members containing different organism compositions.
Table 36. Qualitative Site to Site Results
Target Concentration/Sample
Percent Agreement with Expected Results
Site 1 Site 2 Site 3 All
BacterialVaginosis
True Negative 100192/192
100192/192
100192/192
100576/576
BV Negative 10032/32
10032/32
10032/32
10096/96
Low BV Positivea 10064/64
96.962/64
10064/64
99.0190/192
Moderate Positivea 10064/64
98.463/64
10064/64
99.5191/192
Trichomonas vaginalis
True Negative 100160/160
100160/160
100160/160
100480/480
Low Positive 10032/32
10032/32
10032/32
10096/96
Moderate Positive 10032/32
10032/32
10032/32
10096/96
Candida albicans
True Negative 98.8158/160
98.8158/160
98.1157/160
98.5473/480
Low Positive 10032/32
10032/32
96.931/32
99.095/96
Moderate Positive 10032/32
10032/32
10032/32
10096/96
Candida glabrataTrue Negative 100
160/160100
160/160100
160/160100
480/480
Low Positive 10032/32
96.931/32
10032/32
99.095/96
Candida kruseiTrue Negative 99.4
159/16099.4
159/160100
160/16099.6
478/480
Low Positive 10032/32
10032/32
96.931/32
99.095/96
a Performance includes combined results from replicates of two panel members containing different organism compositions.
To evaluate the BD MAX Vaginal Panel lot-to-lot reproducibility, testing was performed at 1 site with 3 lots over 8 days. At the testing site, 2 operators performed 2 runs on alternate days, for a total of 48 runs. The overall lot-to-lot reproducibility percent agreements ranged from 99.2 to 100%. Reproducibility results are reported in Table 38. The qualitative and quantitative reproducibility results across lots and by target are presented in Tables 39–40. Second Derivative Peak Abscissa (SDPA), an internal criterion used to determine a final assay result, was selected as a means of assessing quantitative assay reproducibility. Mean SDPA values with variance components (SD and% CV) are shown in Table 40.
Table 38. Qualitative Reproducibility Study Results Summary - Lot to Lot
a The expected assay results were deemed to be negative.b Performance includes combined results from replicates of two panel members containing different organism compositions.
34
Table 39. Qualitative Lot-to-Lot Results
Target Concentration/Sample
Percent Agreement with Expected ResultsLot 1 Lot 2 Lot 3 All
BacterialVaginosis
True Negative 100192/192
100192/192
100192/192
100576/576
BV Negative 10032/32
10032/32
10032/32
10096/96
Low BV Positivea 100 64/64
10064/64
10064/64
100192/192
Moderate Positivea 10064/64
10064/64
10064/64
100192/192
Trichomonas vaginalis
True Negative 100160/160
100160/160
100160/160
100480/480
Low Positive 10032/32
10032/32
10032/32
10096/96
Moderate Positive 10032/32
10032/32
10032/32
10096/96
Candida albicans
True Negative 98.8158/160
99.4159/160
99.4159/160
99.2476/480
Low Positive 10032/32
10032/32
10032/32
10096/96
Moderate Positive 10032/32
10032/32
10032/32
10096/96
Candida glabrataTrue Negative 100
160/160100
160/160100
160/160100
480/480
Low Positive 10032/32
10032/32
10032/32
10096/96
Candida kruseiTrue Negative 99.4
159/160100
160/160100
160/16099.8
479/480
Low Positive 10032/32
10032/32
10032/32
10096/96
a Performance includes combined results from replicates of two panel members containing different organism compositions.
Carry-over/Cross-ContaminationA study was conducted to investigate within-run and between-run carry-over while processing samples with high bacterial loads of BD MAX Vaginal Panel targets. A panel made of one high positive member and one negative member was used. In the high positive panel member, vaginosis markers were represented by a combination of L. jensenii (2.7 x 105 CFU/mL), G. vaginalis (2.0 x 106 CFU/mL), A. vaginae (3.2 x 106 CFU/mL), and BVAB-2 (3.5 x 107 cp/mL) while vaginitis targets were represented by T. vaginalis (8.0 x 103 cells/mL). The negative member did not contain any target analyte. Twelve (12) replicates of the high positive panel member and 12 replicates of the negative panel member were tested in each run by alternating negative and positive samples. Three (3) operators performed 3 consecutive runs across 3 BD MAX instruments for a total of 9 runs containing 24 samples. In the 107 negative samples yielding reportable results, neither false positive bacterial vaginosis nor false positive Trichomonas vaginalis results were obtained.
35
REFERENCES1. Centers for Disease Control and Prevention, Sexually Transmitted Disease Surveillance 2014, Atlanta: U.S., Department of
Health and Human Services; 2015. (http://www.cdc.gov/std/stats14)2. Egan ME, Lipsky MS (2000). Diagnosis of Vaginitis. Am Fam Physician 62 (5): 1095–1104 3. Fredricks DN, Fiedler TL, et al. (2007). “Targeted PCR for detection of vaginal bacteria associated with bacterial vaginosis.”
J Clin Microbiol 45(10): 3270–3276.4. Menard JP, Fenollar F, et al. (2008). “Molecular quantification of Gardnerella vaginalis and Atopobium vaginae loads to predict
bacterial vaginosis.” Clin Infect Dis 47(1): 33–43.5. McCormack WM. “Vulvovaginitis and cervicitis”. In: Mandell GL, Bennett JE, Dolin R, (eds). Principles and Practice of Infectious
Diseases. 7th ed. Philadelphia, PA: Elsevier Churchill Livingstone; 2009:chap 107.6. Shipitsyna E, Roos A, et al. (2013). “Composition of the vaginal microbiota in women of reproductive age-sensitive and specific
molecular diagnosis of bacterial vaginosis is possible?” PLoS One 8(4): e60670.7. Srinivasan S, Hoffman NG, et al. (2012). “Bacterial communities in women with bacterial vaginosis: high resolution phylogenetic
analyses reveal relationships of microbiota to clinical criteria.” PLoS One 7(6): e37818.8. Centers for Disease Control and Prevention. Sexually Transmitted Diseases Treatment Guidelines, 2015. MMWR Recomm Rep
2015;64(No. RR-3):1–138.9. Hainer BL and Gibson MV. (2011). “Vaginitis: Diagnosis and Treatment.” Am Fam Physician 83(7): 807–815. 10. Vermitsky JP, Self MJ, et al. (2008). “Survey of vaginal-flora Candida species isolates from women of different age groups by
use of species-specific PCR detection.” J Clin Microbiol 46(4): 1501–1503.11. Paulitsch A, Weger W, et al. (2006). “A 5-year (2000-2004) epidemiological survey of Candida and non-Candida yeast species
causing vulvovaginal candidiasis in Graz, Austria.” Mycoses 49(6): 471–475.12. Fan SR, Liu XP, et al. (2008). “Clinical characteristics of vulvovaginal candidiasis and antifungal susceptibilities of Candida
species isolates among patients in southern China from 2003 to 2006.” J Obstet Gynaecol Res 34(4): 561–566.13. Richter SS, Galask RP, et al. (2005). “Antifungal susceptibilities of Candida species causing vulvovaginitis and epidemiology of
recurrent cases.” J Clin Microbiol 43(5): 2155–2162.14. Kennedy MA and Sobel JD (2010). “Vulvovaginal Candidiasis Caused by Non-albicans Candida Species: New Insights.” Curr
Infect Dis Rep 12(6): 465–470.15. Mahmoudi Rad M, Zafarghandi A, et al. (2012). “Identification of Candida species associated with vulvovaginal candidiasis by
multiplex PCR.” Infect Dis Obstet Gynecol 2012: 872169, doi:10.1155/2012/872169.16. Fidel Jr. PL, Vazquez JA, et al. (1999). “Candida glabrata: review of epidemiology, pathogenesis, and clinical disease with
comparison to C. albicans.” Clin Microbiol Rev 12(1): 80–96.17. Pfaller MA. (2008). “Candida krusei, a multidrug-resistant opportunistic fungal pathogen: geographic and temporal trends from
the ARTEMIS DISK Antifungal Surveillance Program, 2001 to 2005.” J Clin Microbiol. 46(2):515–21.18. Sobel JD. (2007). “Vulvovaginal candidosis.” Lancet. 369 : 1961–71.19. Linhares IM, Witkin SS, et al. (2001).” Differentiation between women with vulvovaginal symptoms who are positive or negative
for Candida species by culture. “ Infect Dis Obstet Gynecol 9 : 221–225. 20. Shafir SC, Sorvillo FJ, et al. (2009). “Current Issues and Considerations Regarding Trichomoniasis and Human
Immunodeficiency Virus in African-Americans.” Clin Microbiol Rev 22: 37–45.21. Lowe NK, Neal JL, et al. (2009). “Accuracy of the clinical diagnosis of vaginitis compared with a DNA probe laboratory
standard.” Obstet Gynecol. 113(1):89–95. 22. Clinical and Laboratory Standards Institute. Protection of laboratory workers from occupationally acquired infections; Approved
Guideline. Document M29 (Refer to the latest edition).23. Centers for Disease Control and Prevention, and National Institutes of Health. Biosafety in microbiological and biomedical
laboratories. Chosewood L.C. and Wislon D.E. (eds) (2009). HHS Publication No. (CDC) 21–1112.24. BD MAX System User’s Manual (Refer to the latest version) BD Life Sciences, Sparks, MD 21152 USA. 25. Clinical and Laboratory Standards Institute. Molecular Diagnostic Methods for Infectious Diseases; Approved Guideline,
Document MM3 (Refer to the latest edition).26. Clinical and Laboratory Standards Institute. 2006. Approved Guideline, Document C24. Statistical Quality Control for
Quantitative Measurements: Principles and Definitions, 3rd ed., CLSI. Wayne PA (Refer to latest edition).27. Clinical and Laboratory Standards Institute. User Protocol for Evaluation of Qualitative Test Performance; Approved Guideline,
Document EP12 (Refer to the latest edition).28. Marot-Leblond A. et al. (2009). “Efficient diagnosis of vulvovaginal candidiasis by use of a new rapid immunochromatography
test” J Clin Microbiol 47 (12): 3821.29. Tibaldi C. et al. (2009). “Vaginal and endocervical microorganisms in symptomatic and asymptomatic non-pregnant females: risk
factors and rates of occurrence” Clin. Microbiol. Infect., 15 :670–679.30. Ravel J. et al. (2013). “Daily temporal dynamics of vaginal microbiota before, during and after episodes of bacterial vaginosis”
Microbiome 1:29–35.
36
Revision Date Change Summary(06) 2019-02 Added statement to Limitations of the Procedure section.
(07) 2019-10 Modification of clinical and analytical data tables according to ADF revision 3.
(08) 2020-05 Converted printed instructions for use to electronic format and added access information to obtain the document from bd.com/e-labeling. Added additional information to Reagents and Materials section. Updated Warnings and Precautions section. Added detail to Instructions for Use section. Updated Figures 2, 3, and 4. Corrected any reference to the System Error Summary section to refer to the Troubleshooting section. Updated Limitations of the Procedure. Updated Australia and New Zealand Sponsor addresses. Made typographical edits.
US Customers only: For symbol glossary, refer to bd.com/symbols-glossary
Manufacturer/Производител/Výrobce/Fabrikant/Hersteller/Κατασκευαστής/Fabricante/Tootja/Fabricant/Proizvođać/Gyártó/Fabbricante/Атқарушы/제조업체/Gamintojas/Ražotājs/Tilvirker/Producent/Producător/Производитель/Výrobca/Proizvođač/Tillverkare/Üretici/Виробник/生产厂商 Useby/Използвайтедо/Spotřebujtedo/Brugfør/Verwendbarbis/Χρήσηέως/Usarantesde/Kasutadaenne/Datedepéremption/사용 기한 / Upotrijebiti do/Felhasználhatóságdátuma/Usareentro/Дейінпайдалануға/Naudokiteiki/Izlietotlīdz/Houdbaartot/Brukesfor/Stosowaćdo/Prazodevalidade/Aseutilizapânăla/Использоватьдо/Použitedo/Upotrebitido/Användföre/Sonkullanmatarihi/Використатидо\line/使用截止日期YYYY-MM-DD / YYYY-MM (MM = end of month)ГГГГ-ММ-ДД/ГГГГ-ММ(ММ=краянамесеца)RRRR-MM-DD/RRRR-MM(MM=konecměsíce)ÅÅÅÅ-MM-DD / ÅÅÅÅ-MM (MM = slutning af måned)JJJJ-MM-TT / JJJJ-MM (MM = Monatsende)ΕΕΕΕ-MM-HH/ΕΕΕΕ-MM(MM=τέλοςτουμήνα)AAAA-MM-DD / AAAA-MM (MM = fin del mes)AAAA-KK-PP / AAAA-KK (KK = kuu lõpp)AAAA-MM-JJ / AAAA-MM (MM = fin du mois)GGGG-MM-DD / GGGG-MM (MM = kraj mjeseca)ÉÉÉÉ-HH-NN/ÉÉÉÉ-HH(HH=hónaputolsónapja)AAAA-MM-GG / AAAA-MM (MM = fine mese)ЖЖЖЖ-АА-КК/ЖЖЖЖ-АА/(АА=айдыңсоңы)YYYY-MM-DD/YYYY-MM(MM = 월말)MMMM-MM-DD/MMMM-MM(MM=mėnesiopabaiga)GGGG-MM-DD/GGGG-MM(MM=mēnešabeigas)JJJJ-MM-DD / JJJJ-MM (MM = einde maand)ÅÅÅÅ-MM-DD / ÅÅÅÅ-MM (MM = slutten av måneden)RRRR-MM-DD/RRRR-MM(MM=koniecmiesiąca)AAAA-MM-DD / AAAA-MM (MM = fim do mês)AAAA-LL-ZZ/AAAA-LL(LL=sfârşitullunii)ГГГГ-ММ-ДД/ГГГГ-ММ(ММ=конецмесяца)RRRR-MM-DD / RRRR-MM (MM = koniec mesiaca)GGGG-MM-DD / GGGG-MM (MM = kraj meseca)ÅÅÅÅ-MM-DD / ÅÅÅÅ-MM (MM = slutet av månaden)YYYY-AA-GG/YYYY-AA(AA=ayınsonu)РРРР-MM-ДД/РРРР-MM(MM=кінецьмісяця)YYYY-MM-DD / YYYY-MM (MM = 月末)
Catalognumber/Каталоженномер/Katalogovéčíslo/Katalognummer/Αριθμόςκαταλόγου/Númerodecatálogo/Katalooginumber/Numérocatalogue/Kataloškibroj/Katalógusszám/Numerodicatalogo/Каталогнөмірі/카탈로그 번호 / Katalogo / numeris / Kataloga numurs / Catalogus nummer / Numer katalogowy/Numărdecatalog/Номерпокаталогу/Katalógovéčíslo/Kataloškibroj/Katalognumarası/Номерзакаталогом/目录号
AuthorizedRepresentativeintheEuropeanCommunity/ОторизиранпредставителвЕвропейскатаобщност/AutorizovanýzástupceproEvropskémspolečenství/AutoriseretrepræsentantiDeEuropæiskeFællesskaber/AutorisierterVertreterinderEuropäischenGemeinschaft/ΕξουσιοδοτημένοςαντιπρόσωποςστηνΕυρωπαϊκήΚοινότητα/RepresentanteautorizadoenlaComunidadEuropea/VolitatudesindajaEuroopaNõukogus/ReprésentantautorisépourlaCommunautéeuropéenne/AutorizuiranipredstavnikuEuropskojuniji/MeghatalmazottképviselőazEurópaiKözösségben/RappresentanteautorizzatonellaComunitàEuropea/Европақауымдастығындағыуәкілеттіөкіл/유럽 공동체의 위임 대표 /ĮgaliotasisatstovasEuroposBendrijoje/PilnvarotaispārstāvisEiropasKopienā/BevoegdevertegenwoordigerindeEuropeseGemeenschap/AutorisertrepresentantiEU/AutoryzowaneprzedstawicielstwoweWspólnocieEuropejskiej/RepresentanteautorizadonaComunidadeEuropeia/ReprezentantulautorizatpentruComunitateaEuropeană/УполномоченныйпредставительвЕвропейскомсообществе/AutorizovanýzástupcavEurópskomspoločenstve/AutorizovanopredstavništvouEvropskojuniji/AuktoriseradrepresentantiEuropeiskagemenskapen/AvrupaTopluluğuYetkiliTemsilcisi/УповноваженийпредставникукраїнахЄС/欧洲共同体授权代表
InVitroDiagnosticMedicalDevice/Медицинскиуредзадиагностикаинвитро/Lékařskézařízeníurčenéprodiagnostikuinvitro/Invitrodiagnostiskmedicinskanordning/MedizinischesIn-vitro-Diagnostikum/Invitroδιαγνωστικήιατρικήσυσκευή/Dispositivomédicoparadiagnósticoinvitro/Invitrodiagnostikameditsiiniaparatuur/Dispositifmédicaldediagnosticinvitro/MedicinskapomagalazaInVitroDijagnostiku/Invitrodiagnosztikaiorvosieszköz/Dispositivomedicaleperdiagnosticainvitro/Жасандыжағдайдажүргізетінмедициналықдиагностикааспабы/InVitro Diagnostic 의료 기기 / In vitro diagnostikos prietaisas/Medicīnasierīces,kolietoinvitrodiagnostikā/Medischhulpmiddelvoorin-vitrodiagnostiek/Invitrodiagnostiskmedisinskutstyr/Urządzeniemedycznedodiagnostykiinvitro/Dispositivomédicoparadiagnósticoinvitro/Dispozitivmedicalpentrudiagnosticinvitro/Медицинскийприбордлядиагностикиinvitro/Medicínskapomôckanadiagnostikuinvitro/Medicinskiuređajzainvitrodijagnostiku/Medicintekniskproduktförinvitro-diagnostik/İnVitroDiyagnostikTıbbiCihaz/Медичнийпристрійдлядіагностикиinvitro/体外诊断医疗设备
Temperaturelimitation/Температурниограничения/Teplotníomezení/Temperaturbegrænsning/Temperaturbegrenzung/Περιορισμοίθερμοκρασίας/Limitacióndetemperatura/Temperatuuripiirang/Limitesdetempérature/Dozvoljenatemperatura/Hőmérsékletihatár/Limitiditemperatura/Температуранышектеу/온도 제한 /Laikymotemperatūra/Temperatūrasierobežojumi/Temperatuurlimiet/Temperaturbegrensning/Ograniczenietemperatury/Limitesdetemperatura/Limitedetemperatură/Ограничениетемпературы/Ohraničenieteploty/Ograničenjetemperature/Temperaturgräns/Sıcaklıksınırlaması/Обмеженнятемператури/ 温度限制
BatchCode(Lot)/Коднапартидата/Kód(číslo)šarže/Batch-kode(lot)/Batch-Code(Charge)/Κωδικόςπαρτίδας(παρτίδα)/Códigodelote(lote)/Partiikood/Numérodelot/Lot(kod)/Tételszáma(Lot)/Codicebatch(lotto)/Топтамакоды/배치 코드(로트) / Partijos numeris (LOT) / Partijas kods (laidiens) / Lot nummer / Batch-kode (parti) /Kodpartii(seria)/Códigodolote/Coddeserie(Lot)/Кодпартии(лот)/Kódsérie(šarža)/Kodserije/Partinummer(Lot)/PartiKodu(Lot)/Кодпартії/批号（亚批）
Containssufficientfor<n>tests/Съдържаниетоедостатъчноза<n>теста/Dostatečnémnožstvípro<n>testů/Indeholdertilstrækkeligttil<n>tests/Ausreichendfür<n>Tests/Περιέχειεπαρκήποσότηταγια<n>εξετάσεις/Contenidosuficientepara<n>pruebas/Küllaldane<n>testidejaoks/Contenusuffisantpour<n>tests/Sadržajza<n>testova/<n>teszthezelegendő/Contenutosufficienteper<n>test/<п>тесттеріүшінжеткілікті/<n>테스트가 충분히 포함됨 / Pakankamas kiekis atlikti<n>testų/Saturpietiekami<n>pārbaudēm/Inhoudvoldoendevoor“n”testen/Innholdertilstrekkeligtil<n>tester/Zawierailośćwystarczającądo<n>testów/Conteúdosuficientepara<n>testes/Conţinutsuficientpentru<n>teste/Достаточнодля<n>тестов(а)/Obsahvystačína<n>testov/Sadržajdovoljanza<n>testova/Innehållertillräckligtför<n>analyser/<n>testiçinyeterlimalzemeiçerir/Вистачитьдляаналізів:<n>/足够进行 <n>次检测
Donotreuse/Неизползвайтеотново/Nepoužívejteopakovaně/Ikketilgenbrug/Nichtwiederverwenden/Μηνεπαναχρησιμοποιείτε/Noreutilizar/Mittekasutadakorduvalt/Nepasréutiliser/Nekoristitiponovo/Egyszerhasználatos/Nonriutilizzare/Пайдаланбаңыз/재사용 금지 / Tik vienkartiniam naudojimui / Nelietotatkārtoti/Nietopnieuwgebruiken/Kuntilengangsbruk/Niestosowaćpowtórnie/Nãoreutilize/Nurefolosiţi/Неиспользоватьповторно/Nepoužívajteopakovane/Neupotrebljavajteponovo/Fårejåteranvändas/Tekrarkullanmayın/Невикористовуватиповторно/请勿重复使用
Serialnumber/Сериенномер/Sériovéčíslo/Serienummer/Seriennummer/Σειριακόςαριθμός/Nºdeserie/Seerianumber/Numérodesérie/Serijskibroj/Sorozatszám/Numerodiserie/Топтамалықнөмірі/일련 번호 /Serijosnumeris/Sērijasnumurs/Serienummer/Numerseryjny/Númerodesérie/Numărdeserie/Серийныйномер/Serinumarası/Номерсерії/序列号
ForIVDPerformanceevaluationonly/СамозаоценкакачествотонаработанаIVD/PouzeprovyhodnocenívýkonuIVD/KuntilevalueringafIVDydelse/NurfürIVD-Leistungsbewertungszwecke/MόνογιααξιολόγησηαπόδοσηςIVD/Sóloparalaevaluacióndelrendimientoendiagnósticoinvitro/AinultIVDseadmehindamiseks/Réservéàl’évaluationdesperformancesIVD/SamouznanstvenesvrhezaInVitroDijagnostiku/Kizárólaginvitrodiagnosztikához/SolopervalutazionedelleprestazioniIVD/Жасандыжағдайда«пробиркаішінде»,диагностикадатекжұмыстыбағалауүшін/IVD 성능 평가에 대해서만 사용 / Tik IVD prietaisųveikimocharakteristikomstikrinti/VienīgiIVDdarbībasnovērtēšanai/Uitsluitendvoordoeltreffendheidsonderzoek/KunforevalueringavIVD-ytelse/TylkodoocenywydajnościIVD/UsoexclusivoparaavaliaçãodeIVD/NumaipentruevaluareaperformanţeiIVD/Толькодляоценкикачествадиагностикиinvitro/Určenéibanadiagnostikuinvitro/Samozaprocenuučinkauinvitrodijagnostici/Endastförutvärderingavdiagnostiskanvändninginvitro/YalnızcaIVDPerformansdeğerlendirmesiiçin/Тількидляоцінюванняякостідіагностикиinvitro/仅限 IVD 性能评估 For US: “For Investigational Use Only”Lowerlimitoftemperature/Доленлимитнатемпературата/Dolníhraniceteploty/Nedretemperaturgrænse/Temperaturuntergrenze/Κατώτεροόριοθερμοκρασίας/Límiteinferiordetemperatura/Aluminetemperatuuripiir/Limiteinférieuredetempérature/Najnižadozvoljenatemperatura/Alsóhőmérsékletihatár/Limiteinferioreditemperatura/Температураныңтөменгіруқсатшегі/하한 온도 /Žemiausialaikymotemperatūra/Temperatūraszemākārobeža/Laagstetemperatuurlimiet/Nedretemperaturgrense/Dolnagranicatemperatury/Limiteminimodetemperatura/Limităminimădetemperatură/Нижнийпределтемпературы/Spodnáhranicateploty/Donjagranicatemperature/Nedretemperaturgräns/Sıcaklıkaltsınırı/Мінімальнатемпература/温度下限
Positivecontrol/Положителенконтрол/Pozitivníkontrola/Positivkontrol/PositiveKontrolle/Θετικόςμάρτυρας/Controlpositivo/Positiivnekontroll/Contrôlepositif/Pozitivnakontrola/Pozitívkontroll/Controllopositivo/Оңбақылау/ 양성 컨트롤 /Teigiamakontrolė/Pozitīvākontrole/Positievecontrole/Kontroladodatnia/Controlopositivo/Controlpozitiv/Положительныйконтроль/Pozitifkontrol/Позитивнийконтроль/阳性对照试剂
Negativecontrol/Отрицателенконтрол/Negativníkontrola/Negativkontrol/NegativeKontrolle/Αρνητικόςμάρτυρας/Controlnegativo/Negatiivnekontroll/Contrôlenégatif/Negativnakontrola/Negatívkontroll/Controllonegativo/Негативтікбақылау/ 음성 컨트롤 /Neigiamakontrolė/Negatīvākontrole/Negatievecontrole/Kontrolaujemna/Controlonegativo/Controlnegativ/Отрицательныйконтроль/Negatifkontrol/Негативнийконтроль/阴性对照试剂
BiologicalRisks/Биологичнирискове/Biologickárizika/Biologiskfare/Biogefährdung/Βιολογικοίκίνδυνοι/Riesgosbiológicos/Bioloogilisedriskid/Risquesbiologiques/Biološkirizik/Biológiailagveszélyes/Rischiobiologico/Биологиялықтәуекелдер/ 생물학적 위험 /Biologinispavojus/Bioloģiskieriski/Biologischrisico/Biologiskrisiko/Zagrożeniabiologiczne/Perigobiológico/Riscuribiologice/Биологическаяопасность/Biologickériziko/Biološkirizici/Biologiskrisk/BiyolojikRiskler/Біологічнанебезпека/生物学风险
Upperlimitoftemperature/Горенлимитнатемпературата/Horníhraniceteploty/Øvretemperaturgrænse/Temperaturobergrenze/Ανώτεροόριοθερμοκρασίας/Límitesuperiordetemperatura/Üleminetemperatuuripiir/Limitesupérieuredetempérature/Gornjadozvoljenatemperatura/Felsőhőmérsékletihatár/Limitesuperioreditemperatura/Температураныңруқсатетілгенжоғарғышегі/상한 온도 /Aukščiausialaikymotemperatūra/Augšējātemperatūrasrobeža/Hoogstetemperatuurlimiet/Øvretemperaturgrense/Górnagranicatemperatury/Limitemáximodetemperatura/Limitămaximădetemperatură/Верхнийпределтемпературы/Hornáhranicateploty/Gornjagranicatemperature/Övretemperaturgräns/Sıcaklıküstsınırı/Максимальнатемпература/温度上限
Keepdry/Пазетесухо/Skladujtevsuchémprostředí/Opbevarestørt/Trocklagern/Φυλάξτετοστεγνό/Mantenerseco/Hoidakuivas/Conserverausec/Držatinasuhom/Szárazhelyentartandó/Tenereall’asciutto/Құрғақкүйіндеұста/ 건조 상태 유지 /Laikykitesausai/Uzglabātsausu/Drooghouden/Holdestørt/Przechowywaćwstaniesuchym/Manterseco/Aseferideumezeală/Недопускатьпопаданиявлаги/Uchovávajtevsuchu/Držitenasuvommestu/Förvarastorrt/Kurubirşekildemuhafazaedin/Берегтивідвологи/请保持干燥
Collectiontime/Временасъбиране/Časodběru/Opsamlingstidspunkt/Entnahmeuhrzeit/Ώρασυλλογής/Horaderecogida/Kogumisaeg/Heuredeprélèvement/Satiprikupljanja/Mintavételidőpontja/Oradiraccolta/Жинаууақыты/수집 시간 /Paėmimolaikas/Savākšanaslaiks/Verzameltijd/Tidprøvetaking/Godzinapobrania/Horadecolheita/Oracolectării/Времясбора/Dobaodberu/Vremeprikupljanja/Uppsamlingstid/Toplamazamanı/Часзабору/采集时间
Donotuseifpackagedamaged/Неизползвайте,акоопаковкатаеповредена/Nepoužívejte,je-liobalpoškozený/Måikkeanvendeshvisemballagenerbeskadiget/InhalbeschädigterPackungnichtverwenden/Μηχρησιμοποιείτεεάνησυσκευασίαέχειυποστείζημιά./Nousarsielpaqueteestádañado/Mittekasutada,kuipakendonkahjustatud/Nepasl’utilisersil’emballageestendommagé/Nekoristitiakojeoštećenopakiranje/Nehasználja,haacsomagolássérült/Nonusareselaconfezioneèdanneggiata/Егерпакетбұзылғанболса,пайдаланба/ 패키지가 손상된 경우 사용 금지 /Jeipakuotėpažeista,nenaudoti/Nelietot,jaiepakojumsbojāts/Nietgebruikenindiendeverpakkingbeschadigdis/Måikkebrukeshvispakkeerskadet/Nieużywać,jeśliopakowaniejestuszkodzone/Nãousarseaembalagemestiverdanificada/Anusefolosidacăpachetulestedeteriorat/Неиспользоватьприповрежденииупаковки/Nepoužívajte,akjeobalpoškodený/Nekoristiteakojepakovanjeoštećeno/Användejomförpackningenärskadad/Ambalajhasargörmüşsekullanmayın/Невикористовуватизапошкодженоїупаковки/如果包装破损，请勿使用
Keepawayfromheat/Пазетеоттоплина/Nevystavujtepřílišnémuteplu/Måikkeudsættesforvarme/VorWärmeschützen/Κρατήστετομακριάαπότηθερμότητα/Manteneralejadodefuentesdecalor/Hoidaeemalvalgusest/Protégerdelachaleur/Držatidaljeodizvoratopline/Óvjaamelegtől/Tenerelontanodalcalore/Салқынжердесақта/ 열을 피해야 함 /Laikytiatokiaunuošilumosšaltinių/Sargātnokarstuma/Beschermentegenwarmte/Måikkeutsettesforvarme/Przechowywaćzdalaodźródełciepła/Manteraoabrigodocalor/Aseferidecăldură/Ненагревать/Uchovávajtemimozdrojatepla/Držitedaljeodtoplote/Fårejutsättasförvärme/Isıdanuzaktutun/Берегтивіддіїтепла/请远离热源
Keepawayfromlight/Пазетеотсветлина/Nevystavujtesvětlu/Måikkeudsættesforlys/VorLichtschützen/Κρατήστετομακριάαπότοφως/Manteneralejadodelaluz/Hoidaeemalvalgusest/Conserveràl’abridelalumière/Držatidaljeodsvjetla/Fénynemérheti/Tenerealriparodallaluce/Қараңғыланғанжердеұста/ 빛을 피해야 함 /Laikytiatokiaunuošilumosšaltinių/Sargātnogaismas/Nietblootstellenaanzonlicht/Måikkeutsettesforlys/Przechowywaćzdalaodźródełświatła/Manteraoabrigodaluz/Feriţidelumină/Хранитьвтемноте/Uchovávajtemimodosahusvetla/Držitedaljeodsvetlosti/Fårejutsättasförljus/Işıktanuzaktutun/Берегтивіддіїсвітла/请远离光线
Hydrogengasgenerated/Образуваневодородгаз/Možnostúnikuplynnéhovodíku/Frembringerhydrogengas/Wasserstoffgaserzeugt/Δημιουργίααερίουυδρογόνου/Produccióndegasdehidrógeno/Vesinikgaasitekitatud/Produitdel’hydrogènegazeux/Sadržihydrogenvodik /Hidrogéngáztfejleszt/Produzionedigasidrogeno/Газтектессутегіпайдаболды/ 수소 가스 생성됨/Išskiriavandeniliodujas/Rodasūdeņradis/Waterstofgasgegenereerd/Hydrogengassgenerert/Powodujepowstawaniewodoru/Produçãodegásdehidrogénio/Generaregazdehidrogen/Выделениеводорода/Vyrobenépoužitímvodíka/Oslobađasevodonik/Genereradvätgas/Açığaçıkanhidrojengazı/Реакціязвиділеннямводню/会产生氢气
PatientIDnumber/ИДномернапациента/IDpacienta/PatientensID-nummer/Patienten-ID/Αριθμόςαναγνώρισηςασθενούς/NúmerodeIDdelpaciente/PatsiendiID/Nod’identificationdupatient/Identifikacijskibrojpacijenta/Betegazonosítószáma/NumeroIDpaziente/Пациенттіңидентификациялықнөмірі/ 환자 ID 번호 /Pacientoidentifikavimonumeris/PacientaIDnumurs/Identificatienummervandepatiënt/PasientensID-nummer/NumerIDpacjenta/NúmerodaIDdodoente/NumărIDpacient/Идентификационныйномерпациента/Identifikačnéčíslopacienta/IDbrojpacijenta/Patientnummer/Hastakimliknumarası/Ідентифікаторпацієнта/患者标识号
Fragile,HandlewithCare/Чупливо,Работетеснеобходимотовнимание./Křehké.Přimanipulacipostupujteopatrně./Forsigtig,kangåistykker./Zerbrechlich,vorsichtighandhaben./Εύθραυστο.Χειριστείτετομεπροσοχή./Frágil.Manipularconcuidado./Õrn,käsitsegeettevaatlikult./Fragile.Manipuleravecprécaution./Lomljivo,rukujtepažljivo./Törékeny!Óvatosankezelendő./Fragile,maneggiareconcura./Сынғыш,абайлаппайдаланыңыз./조심 깨지기 쉬운 처리 / Trapu, elkitėsatsargiai./Trausls;rīkotiesuzmanīgi/Breekbaar,voorzichtigbehandelen./Ømtålig,håndterforsiktig./Kruchazawartość,przenosićostrożnie./Frágil,ManuseiecomCuidado./Fragil,manipulaţicuatenţie./Хрупкое!Обращатьсясосторожностью./Krehké,vyžadujesaopatrnámanipulácia./Lomljivo-rukujtepažljivo./Bräckligt.Hanteraförsiktigt./KolayKırılır,DikkatliTaşıyın./Тендітна,звертатисязобережністю/易碎，小心轻放
This only applies to US: “Caution: Federal Law restricts this device to sale by or on the order of a licensed practitioner.” / S’applique uniquement aux États-Unis: “Caution: Federal Law restricts this device to sale by or on the order of a licensed practitioner.” / Vale solo per gli Stati Uniti: “Caution: Federal Law restricts this device to sale by or on the order of a licensed practitioner.” / Gilt nur für die USA: “Caution: Federal Law restricts this device to sale by or on the order of a licensed practitioner.”/SóloseaplicaalosEE.UU.:“Caution:FederalLawrestrictsthisdevicetosalebyorontheorderofalicensedpractitioner.”
Technical Information: In the United States contact BD Technical Service and Support at 1.800.638.8663 or bd.com.
GeneOhm Sciences Canada, Inc. 2555 Boul. du Parc Technologique Québec,QC,G1P4S5,Canada
Benex Limited Pottery Road, Dun Laoghaire Co. Dublin, Ireland
Australian Sponsor: Becton Dickinson Pty Ltd. 66 Waterloo Road Macquarie Park NSW 2113 Australia
New Zealand Sponsor: Becton Dickinson Limited 14B George Bourke Drive Mt. Wellington Auckland 1060 New Zealand
CLSI document follows and is not to be printed or included in
print files to vendor. NOTES: The CLSI document is to be updated at the same time as the package insert and should be released as a separate PDF document to the web. The package insert should also be released as a separate PDF document to the web. The package insert and CLSI document should be released as one document to SAP. Final art provided to Suzhou/print vendor should not include this page or the CLSI portion of this document. Proofreader: please proof the CLSI document along with the package insert.
Procedure BD MAX™ System for BD MAX™ Vaginal Panel
An automated qualitative in vitro diagnostic test for the direct detection of DNA targets from bacteria associated with bacterial vaginosis (qualitative results reported based on detection and quantitation of targeted organism markers), Candida species associated with vulvovaginal candidiasis, and Trichomonas vaginalis from vaginal swabs in patients who are symptomatic for vaginitis/vaginosis.
Prepared by Date Adopted
Supersedes Procedure #
Review Date Revision Date
Signature
Distributed to # of Copies
Distributed to # of
Copies
Any modifications to this document are the sole responsibility of the facility. This “Sample Procedure” is not intended as a substitute for your facility procedure manual, instrument manual, or reagent labeling/package insert. This “Sample Procedure” is intended as a model for use by your facility to meet the needs of your laboratory.
P0223(08)
1
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
INDICATIONS FOR USEThe BD MAX™ Vaginal Panel performed on the BD MAX System is an automated qualitative in vitro diagnostic test for the direct detection of DNA targets from bacteria associated with bacterial vaginosis (qualitative results reported based on detection and quantitation of targeted organism markers), Candida species associated with vulvovaginal candidiasis, and Trichomonas vaginalis from vaginal swabs in patients who are symptomatic for vaginitis/vaginosis. The test utilizes real-time polymerase chain reaction (PCR) for the amplification of specific DNA targets and utilizes fluorogenic target-specific hybridization probes to detect and differentiate DNA from:• Bacterial vaginosis markers (Individual markers not reported)
○ Lactobacillus spp (L. crispatus and L. jensenii)○ Gardnerella vaginalis○ Atopobium vaginae○ BacterialVaginosisAssociatedBacteria-2(BVAB-2)○ Megasphaera-1
• Candida spp (C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis)• Candida glabrata• Candida krusei• Trichomonas vaginalisThe BD MAX Vaginal Panel is intended to aid in the diagnosis of vaginal infections in women with a clinical presentation consistent with bacterial vaginosis, vulvovaginal candidiasis and trichomoniasis.
SUMMARY AND EXPLANATION OF THE PROCEDUREVaginitis, one of the most common gynecological problems in clinical medicine, accounts for millions of office visits each year.1,2 The three main infectious causes of vaginitis are bacterial vaginosis (BV), yeast vaginitis (candidiasis) and T. vaginalis vaginitis (trichomoniasis).2
Bacterial vaginosis is the most common infectious vaginal complaint, and accounts for 40 to 50 percent of cases in women of childbearing age.2 Hydrogen-peroxide producing Lactobacillus species are important members of the normal vaginal flora and are generally known to decrease in patients suffering from vaginosis. The decrease of these bacteria rather than their colonization of the vagina is a recognized marker of vaginosis.3,4 In opposition to this decrease in Lactobacillus species; there is an increase in well-known species of anaerobes such as Atopobium vaginae and Gardnerella vaginalis.4 Colonization also occurs with non-cultivableanaerobeorganismssuchasBVAB-2andMegasphaera-1.3 Additional microorganisms including Prevotella, Lachnospira, Sneathia, Mobiluncus, Mycoplasma hominis, and Ureaplasma spp. have also been found in women with bacterial vaginosis.5 However, these organisms are less associated with BV due to their relatively low prevalence, sensitivity and/or specificity.3,6,7 Bacterial vaginosis is associated with a high incidence of endometritis and pelvic inflammatory disease following abortion and gynecologic procedures in the general population. Bacterial vaginosis has been also associated with late-term miscarriages, premature rupture of membranes, and preterm birth, and has been strongly linked with an increased risk of human immunodeficiency virus (HIV) and other sexually transmitted diseases.8,9 Although it is known that asymptomatic women may have microbial flora consistent with bacterial vaginosis, treatment is generally not indicated by current guidelines for these patients. Seventeen (17) to 39 percent of women symptomatic for vaginitis are diagnosed with vulvovaginal candidiasis.9 Multiple epidemiologic studies have indicated that Candida albicans is responsible for 65 to 90% of vulvovaginal candidiasis cases10,11,12,13,14,15 and that non-Candida albicans species may be responsible for up to 30% of vulvovaginal candidiasis episodes.13,14,15 Among these species, the most frequently reported in literature are C. glabrata, C. parapsilosis and C. tropicalis. C. glabrata and C. krusei are the main Candida species resistant to fluconazole-based antifungal therapy.8,16,17 Vulvovaginalcandidiasis is usually associated with high Candida loads but symptoms may be associated with lower loads in women predisposedby factors altering the vaginal milieu.18,19 Complications of vulvovaginal candidiasis are rare. Chorioamnionitis in pregnancy andvulvar vestibulitis syndrome have been reported.2 Women may also be colonized with Candida spp. and be asymptomatic.Trichomoniasis, caused by Trichomonas vaginalis, is one of the most common sexually transmitted infections worldwide with over 170 million cases per year.20 Four (4) to 35% of women symptomatic for vaginitis are diagnosed with trichomoniasis.9 This disease is associated with several adverse health outcomes, such as preterm birth, delivery of a low-birth weight infant, and facilitation of sexual transmission of Human Immunodeficiency Virus (HIV).1
Physicians traditionally diagnose vaginitis using the combination of symptoms, physical examination, pH of vaginal fluid, microscopy, and the whiff test. When combined, these tests display a sensitivity and specificity of 81 and 70% respectively for bacterial vaginosis; 84 and 85% for vulvovaginal candidiasis; and 85 and 100% for trichomoniasis when compared with a molecular assay.9,21
2
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
The BD MAX Vaginal Panel is designed for use with the BD MAX UVE Specimen Collection kit. Samples are transported to the testing laboratory in BD MAX UVE Sample Buffer Tubes. The Sample Buffer Tubes are vortexed to release cells from the swab into the buffer. The Sample Buffer Tubes, Unitized Reagent Strips and PCR Cartridges are loaded on the BD MAX System. No further operator intervention is necessary and the following automated procedures occur. The cells are lysed, and DNA is extracted, captured and concentrated on magnetic beads. After an elution step, DNA is added to reagents containing specific primers and probes used to amplify and detect the genetic targets, if present. Upon amplification, signal detection and interpretation are performed automatically by the BD MAX System using real-time PCR. Each Extraction Tube includes a Sample Processing Control, which monitors the integrity of the reagents as well as the process steps involved in DNA extraction, amplification and detection, and checks for the presence of potential assay inhibitors.
PRINCIPLES OF THE PROCEDUREA combination of lytic and extraction reagents is used to perform cell lysis and DNA extraction. Nucleic acids released from the target organisms are captured on magnetic affinity beads. The beads, together with the bound nucleic acids, are washed and the nucleic acids are eluted by a combination of heat and pH variation. Eluted DNA is neutralized and transferred to the Master Mix Tubes to rehydrate the PCR reagents. After reconstitution, the BD MAX System dispenses a fixed volume of PCR-ready solution containing extracted nucleic acids into the PCR Cartridge. Microvalves in the cartridge are sealed by the system prior to initiating PCR in order to contain the amplification mixture and thus prevent evaporation and contamination. The amplified DNA targets are detected using hydrolysis (TaqMan®) probes, labeled at one end with a fluorescent reporter dye (fluorophore), and at the other end, with a quencher moiety. Probes labeled with different fluorophores are used to detect the target analytes in different optical channels of the BD MAX System. When the probes are in their native state, the fluorescence of the fluorophore is quenched due to its proximity to the quencher. However, in the presence of target DNA, the probes hybridize to their complementary sequences and are hydrolyzed by the 5’–3’ exonuclease activity of the DNA polymerase as it synthesizes the nascent strand along the DNA template. As a result, the fluorophores are separated from the quencher molecules and fluorescence is emitted. The amount of fluorescence detected in the optical channels used for the BD MAX Vaginal Panel is directly proportional to the quantity of the corresponding probe that is hydrolyzed. The BD MAX System monitors these signals at each cycle of the PCR and interprets the data at the end of the reaction to provide qualitative test results for each analyte of the vaginitis targets (i.e., positive or negative) and positive or negative BV results are obtained from the combination of bacterial vaginosis marker signals.
WARNINGS AND PRECAUTIONSThe BD MAX Vaginal Panel is for in vitro diagnostic use.• If patient-collected vaginal swabs are not directly transferred into a BD MAX UVE Sample Buffer Tube, they must be transferred
within2hoursofcollectionwhenkeptat2–30°C.• Do not pre-warm samples prior to using the BD MAX Vaginal Panel.• Do not use expired reagents and/or materials.• Do not use the kit if the label that seals the outer box is broken upon arrival.• Do not use reagents if the protective pouches are open or broken upon arrival.• Do not use reagents if desiccant is not present or is broken inside reagent pouches.• Do not remove desiccant from reagent pouches.• Close protective pouches of reagents promptly with the zip seal after each use. Remove any excess air in the pouches prior
to sealing.• Protect reagents against heat and humidity. Prolonged exposure to humidity may affect product performance.• Do not use reagents if the foil has been broken or damaged.• Do not mix reagents from different pouches and/or kits and/or lots.• Do not interchange or reuse caps, as contamination may occur and compromise test results.• CheckUnitizedReagentStripsforproperliquidfills(ensurethattheliquidsareatthebottomofthetubes)(refertoFigure2).• CheckUnitizedReagentStripstoensurethatallpipettetipsarepresent(refertoFigure2).• Proceed with caution when using chemical solutions as Master Mix and Extraction Tube, barcode readability may be altered.• Good laboratory technique is essential to the proper performance of this assay. Due to the high analytical sensitivity of this test,
extreme care should be taken to preserve the purity of all materials and reagents.• In cases where other PCR tests are conducted in the same general area of the laboratory, care must be taken to ensure that
the BD MAX Vaginal Panel, any additional reagents required for testing, and the BD MAX System are not contaminated. Avoidmicrobial and deoxyribonuclease (DNase) contamination of reagents at all times. Gloves must be changed before manipulatingreagents and cartridges.
• To avoid contamination of the environment by amplicons, do not break apart the BD MAX PCR Cartridges after use. The sealsof the BD MAX PCR Cartridges are designed to prevent contamination.
• The laboratory should routinely perform environmental monitoring to minimize the risk of cross-contamination.• Performing the BD MAX Vaginal Panel outside the recommended time ranges can produce invalid results. Assays not
performed within the specified time ranges should be repeated with a new specimen.• Additional controls may be tested according to guidelines or requirements of local, state, provincial and/or federal regulations or
accrediting organizations.• Always handle specimens as if they are infectious and in accordance with safe laboratory procedures such as those described
intheCLSIDocumentM2922 and in Biosafety in Microbiological and Biomedical Laboratories.23
• Wear protective clothing and disposable gloves while handling all reagents.• Wash hands thoroughly after performing the test.• Do not pipette by mouth.• Do not smoke, drink, chew or eat in areas where specimens or kit reagents are being handled.• Dispose of unused reagents and waste in accordance with local, state, provincial and/or federal regulations.• Consult the BD MAX System User’s Manual24 for additional warnings, precautions and procedures.
STORAGE AND STABILITYSpecimen StabilityCollected specimens should be transferred to the BD MAX UVE Sample Buffer Tubes as prescribed in the Specimen Collection kitpackage insert.SpecimensinBDMAXUVESampleBufferTubescanbestoredforamaximumof8daysat2–30°Corforamaximumof14daysat2–8°Cbeforetesting.Kit Components StorageBDMAXVaginalPanelcomponentsarestableat2–25°Cthroughthestatedexpirationdate.Donotuseexpiredcomponents.BD MAX Vaginal Panel Master Mixes and Extraction Tubes are provided in sealed pouches. To protect product from humidity, immediatelyre-sealpouchesafteropening.Reagenttubesarestableforupto14daysat2–25°Cafterinitialopeningandre-sealing of the pouch.UnreconstitutedBDMAXMasterMixesandExtractionTubesarestableforupto24hoursat2–25°Cafterbeingremovedfromtheirprotective pouch.
4
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
INSTRUCTIONS FOR USESpecimen Transport/PreparationNOTE: Wear gloves when handling the BD MAX UVE Specimen Collection kit components and specimens. If gloves come in contact with the specimen, immediately change them to prevent contamination of other specimens.Specimens should be collected using the BD MAX UVE Specimen Collection kit and following the BD MAX UVE Specimen Collection kit instructions. For specimens not yet transferred into the BD MAX UVE Sample Buffer Tube, prepare the samples for testing following the instructions below. Transfer of Vaginal Swab Specimens to the BD MAX UVE Sample Buffer Tube:NOTE: Swabs must be transferred from the swab sheath to the BD MAX UVE Sample Buffer Tube directly (preferred) or within 2 hours of collection when kept at 2–30 °C.NOTE: Refer to Figure 1 for a summary of the swab specimen transfer workflow.1. Uncap the BD MAX UVE Sample Buffer Tube and fully insert the swab into the tube so that the tip is at the bottom.2. Grasping the swab by the cap, carefully break the swab shaft at the score mark. Use caution to avoid splashing or
contamination of the tube contents.3. Tighten the cap securely on the BD MAX UVE Sample Buffer Tube. In the event that the swab shaft is too long to allow closing
the tube securely, request the collection of a new specimen using a new swab.4. Label the BD MAX UVE Sample Buffer Tube with patient information and date/time collected.
NOTE: Be careful not to obscure the barcodes on the tube.5. Proceed directly with Specimen Preparation.
1. Fully insert the swab into the tube so that the tip is at the bottom.
2. Carefully break the shaft at the score mark.
3. Tightly re-cap the tube. 4. Label tube with patient information.
Figure 1: Transfer of Swab Specimens to the BD MAX UVE Sample Buffer Tube
5
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
Specimen Preparation NOTE: One (1) Septum Cap is required for each specimen and each External Control to be tested.1. Place BD MAX UVE Sample Buffer Tubes in a NALGENE™ Cryogenic Vial Holder and vortex at maximum speed for 1 minute
with the Multi-Tube Vortexer. If the run cannot be started within 4 hours of this vortexing step, vortex again at maximumspeed for 1 minute with the Multi-Tube Vortexer before starting the run.
2. Uncap the BD MAX UVE Sample Buffer Tube and gently press the swab against the side of the tube to remove excess fluid.NOTE: If a swab is not captured by the cap, do not attempt to remove it from the BD MAX UVE Sample Buffer Tube. The swabmay remain in the BD MAX UVE Sample Buffer Tube for the whole PCR run.
3. Withdraw the swab/cap combination from the tube and discard in a biomedical waste container. Please note that microbialloads from vaginal specimens can be very high, precautions should be taken to avoid environmental contamination during thisparticular step of the Specimen Preparation.
4. Recap the tube with a blue septum cap.5. Proceed directly with BD MAX System Operation. Do not pre-warm tubes prior to using the BD MAX Vaginal Panel.BD MAX System OperationNOTE: Refer to the BD MAX System User’s Manual24 for detailed instructions (Operation section).NOTE: Testing of the BD MAX Vaginal Panel must be performed within 4 hours after the vortexing step above (refer to Specimen Preparation, Steps 1–3). If retesting is necessary, re-vortex sample(s).NOTE: One (1) Vaginitis Master Mix (C4), one (1) Vaginosis Master Mix (C5), one (1) Extraction Tube (C6), and one (1) BD MAX Vaginal Panel Unitized Reagent Strip are required for each specimen and each External Control to be tested.1. Power on the BD MAX System (if not already done) and log in by entering <user name>and <password>.2. Gloves must be changed before manipulating reagents and cartridges.3. Remove the required number of Unitized Reagent Strips from the BD MAX Vaginal Panel kit. Gently tap each Unitized Reagent
Strip onto a hard surface to ensure that all liquids are at the bottom of the tubes.4. Remove the required number of Extraction Tube(s) and Master Mix Tube(s) from their protective pouches. Remove excess air,
and close pouches with the zip seal.5. For each sample to be tested, place 1 Unitized Reagent Strip on the BD MAX System Rack, starting with Position 1 of Rack A.6. Snap1ExtractionTube(whitefoil)intoeachUnitizedReagentStripinPosition1asshowninFigure2.7. Snap1VaginosisMasterMixTube(greenfoil)intoeachUnitizedReagentStripinPosition2asshowninFigure2.8. Snap1VaginitisMasterMixTube(bluefoil)intoeachUnitizedReagentStripinPosition4asshowninFigure2.
Figure 2: Snap BD MAX Vaginal Extraction Tubes and Master Mix into Unitized Reagent Strips
9. Click on the Run icon, then Inventory. Enter the BD MAX Vaginal Panel kit lot number (for lot traceability) by either scanning thebarcode with the scanner or by manual entry.NOTE: Repeat step 9 each time a new kit lot is used.
10. Navigate to the Worklist. Using the pull down menu select <BD MAX Vaginal 46>.11. Enter the Sample Buffer Tube ID, Patient ID and Accession Number (if applicable) into the Worklist, either by scanning the
barcode with the scanner or by manual entry.12. Select the appropriate kit lot number (found on the outer box) from the pull down menu.13. Repeatsteps10to12forallremainingSampleBufferTubes.
6
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
14. Place the Sample Buffer Tubes in the BD MAX System Rack(s) corresponding to the Unitized Reagent Strips assembled in steps 5 to 8.
15. Place the required number of BD MAX PCR Cartridge(s) into the BD MAX System (refer to Figure 3). • Eachcartridgeaccommodates12samplestestedagainsttheVaginosisandtheVaginitispanelsforatotalof24PCR
reactions per cartridge.• The BD MAX System will automatically select the position and row on the BD MAX PCR Cartridge for each run. BD MAX
PCR Cartridges may be used multiple times until all lanes have been utilized.• TomaximizeuseofBDMAXPCRCartridges,using2000SampleMode,selectRunWizardundertheWorklisttabforlane
assignments.• Consult the BD MAX System User’s Manual24 for more details.
Figure 3: Load PCR Cartridges
16. Load rack(s) onto the BD MAX System (refer to Figure 4).
Side A Side BFigure 4: Load Rack(s) onto BD MAX System
17. Close the BD MAX System lid and click <Start>to begin processing.18. At the end of the run, check results immediately or store Sample Buffer Tubes according to the temperatures and times stated
in “Repeat Test Procedure” (below) until the results are checked.NOTE: If a septum cap was damaged during the run, replace it with a new one before storing the sample.NOTE: After the end of the run, Sample Buffer Tubes containing samples can be stored:
for a maximum of 5 hours at 2–30 °C or for a maximum of 120 hours at 2–8 °C.
When an Indeterminate (IND), Unresolved (UNR), or Incomplete (INC) result is obtained, or when an External Control failure occurs, a repeat test from the prepared Sample Buffer Tube must be performed (see “Repeat Test Procedure”). If an External Control fails, repeat testing of all specimens using freshly prepared External Controls (see “Quality Control”).
7
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
QUALITY CONTROLQuality control procedures monitor the performance of the assay. Laboratories must establish the number, type and frequency of testing control materials according to guidelines or requirements of local, provincial, state, federal and/or country regulations or accreditation organizations. For general Quality Control guidance, the user may wish to refer to Clinical Laboratory Standards Institute documents MM3, C24andEP12.25,26,27
1. External Control materials are not provided by BD. External Positive and Negative Controls are not used by the BD MAXSystem software for the purpose of sample test result interpretation. External Controls are treated as if they were patientsamples. BD MAX UVE Sample Buffer Tubes are needed to prepare External Controls. (Refer to the table in the ResultsInterpretation section for the interpretation of External Control assay results.)
2. One External Positive Control and one External Negative Control should be run at least daily until adequate process validationis achieved on the BD MAX System in each laboratory setting. Each target control strain should be tested alternately. Reducedfrequency of control testing should be in accordance with applicable regulations.
3. The External Positive Control is intended to monitor for substantial reagent failure. The External Negative Control is intended todetect reagent or environmental contamination (or carry-over) by target nucleic acids.
4. Control strains should be tested according to guidelines or requirements of local, state and/or federal regulations oraccreditation organizations in order to monitor the effectiveness of the entire analytical process.
5. Various types of External Controls are recommended to allow the user to select the most appropriate for their laboratory qualitycontrol program.a. External Negative Control: A suspension of commercially available control material [e.g., Lactobacillus iners (ATCC® 55195)]
in a BD MAX UVE Sample Buffer Tube or a previously characterized sample known to be negative. BD recommendsthat the External Negative Control be prepared prior to the External Positive Control in order to reduce the potential forcontamination as a result of control preparation.
b. External Positive Control: A suspension of the commercially available control materials listed below in a BD MAX UVESample Buffer Tube (Table 1) or a previously characterized sample known to be positive.
Table 1: Commercially Available Materials for External Controls
a A mixture of Gardnerella vaginalis and Atopobium vaginae
If control organisms cultures are used:(1) Prepare starting cultures of control organisms as described in the External Control preparation table below.(2) ForallcontrolsotherthanTrichomonas vaginalis, prepare 0.5 McFarland cell suspensions in phosphate buffered saline
(PBS) from fresh culture.(3) PerformserialdilutionslistedinTable2usingPBStoobtainthefinaldilutionrecommendedforeachorganism.(4) For BV Positive Control, mix an equal volume of the 0.5 McFarland of Gardnerella vaginalis ATCC 14018 and of the
0.5 McFarland of Atopobium vaginae ATCC BAA-55. Vortex the mix 5–10 sec at maximum speed. No additional mixingis necessary for other controls.
(6) Place BD MAX UVE Sample Buffer Tubes in a NALGENE Cryogenic Vial Holder and vortex at maximum speed for1 minute with the Multi‑Tube Vortexer. If the run cannot be started within 4 hours of this vortexing step, vortexagain at maximum speed for 1 minute with the Multi-Tube Vortexer before starting the run.
(7) Process the External Control as if it is a patient sample according to the procedure indicated in the BD MAX SystemOperation section.
8
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
Table 2: External Controls Preparation
Controls organism Starting Culture Suspension preparation in PBS Final Dilution
Trichomonas vaginalis ATCC 30001 Use ATCC stock From ATCC stock 1/1000a
Candida albicans ATCC10231 Fresh culture on Sabouraud plate 0.5 McF in PBS 1/20b
Candida glabrataATCC2001 Fresh culture on Sabouraud plate 0.5 McF in PBS 1/100c
Candida krusei ATCC6258 Fresh culture on Sabouraud plate 0.5 McF in PBS 1/20c
Lactobacillus iners ATCC 55195 Fresh culture on anaerobic blood agar plate 0.5 McF in PBS No dilutionc
Gardnerella vaginalis ATCC 14018 Fresh culture on anaerobic blood agar plate 0.5 McF in PBS No dilution: combine an equal volume of each 0.5 McF
suspensioncAtopobium vaginae ATCC BAA-55 Fresh culture on anaerobic blood agar plate 0.5 McF in PBS
a An entire vial of Trichomonas vaginalisATCC30001canbepreparedtothefinaldilution,dividedinto200μLaliquots,frozenandusedforroutine testing.
b Fresh preparation must always be used for Candida albicans ATCC10231.c Suspensioncanbepreparedtothefinaldilution,dividedinto200μLaliquots,frozenandusedforroutinetesting.
6. External Controls should yield the expected results. a. An External Negative Control that yields a positive test result is indicative of a specimen handling and/or contamination
event. Review the specimen handling technique to avoid mix-up and/or contamination. b. An External Positive Control that yields a negative result is indicative of a specimen handling/ preparation problem. Review
the specimen handling/preparation technique.c. An External Control that yields an Unresolved, Indeterminate or Incomplete test result is indicative of a reagent or a
BD MAX System failure. Check the BD MAX System monitor for any error messages. Refer to the Troubleshooting section of the BD MAX System User’s Manual24 for interpretation of warning and error codes. If the problem persists, use reagents from an unopened pouch or use a new assay kit.
7. Each Extraction Tube contains a Sample Processing Control which is a plasmid containing a synthetic target DNA sequence. The Sample Processing Control monitors the efficiency of DNA capture, washing and elution during the sample processing steps, as well as the efficiency of DNA amplification and detection during PCR analysis. If the Sample Processing Control result fails to meet the acceptance criteria, the result of the specimen will be reported as Unresolved for this Master Mix. An Unresolved result is indicative of specimen-associated inhibition or reagent failure. Repeat any specimen reported as Unresolved according to the Repeat Test Procedure section below.
9
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
RESULTS INTERPRETATIONResults are available on the Results tab in the Results window on the BD MAX System monitor. The BD MAX System software automatically interprets test results. A test result may be called NEG (negative), POS (positive) or UNR (unresolved) based on the amplification status of the target and of the Sample Processing Control. IND (indeterminate) or INC (incomplete) results are due to BD MAX System failure. Results interpretation is detailed in Table 3.
Table 3: BD MAX Vaginal Panel Result Interpretation
Assay Result(Vaginitis Master Mix) Interpretation of Results
TV POS Trichomonas vaginalis DNA DetectedTV NEG No Trichomonas vaginalis detected
Cgla POS Candida glabrata DNA DetectedCgla NEG No Candida glabrata detectedCkru POS Candida krusei DNA DetectedCkru NEG No Candida krusei detected
UNR Unresolved – inhibitory sample or reagent failure; No target detected and no amplification of Sample Processing Control
IND Indeterminate result due to BD MAX System failure (with Warning or Error Codes)a
INC Incomplete Run (with Warning or Error Codes)a
Assay Result(Vaginosis Master Mix) Interpretation of Results
BV POS
Vaginosis Panel DNA detected. Detection of marker combinations related to bacterial vaginosis: Gardnerella vaginalis and/or
L. crispatus and/or L. jensenii and/or Atopobium vaginae and/or
BVAB-2and/orMegasphaera-1
BV NEG Detection of marker combinations related to normal vaginal flora
UNR Unresolved – inhibitory sample or reagent failure; No target detected and no amplification of Sample Processing Control
IND Indeterminate result due to BD MAX System failure (with Warning or Error Codes)a
INC Incomplete Run (with Warning or Error Codes)a
a Refer to the Troubleshooting section of the BD MAX System User’s Manual24 for interpretation of warning and error codes.
REPEAT TEST PROCEDURENOTE: Sufficient volume is available for one repeat test from the Sample Buffer Tube. For Sample Buffer Tubes stored at 2–30 °C, retesting must be initiated within 5 hours of the end of the run. Alternatively, for Sample Buffer Tubes stored at 2–8 °C, retesting may be initiated within 120 hours (5 days) of the end of the run. NOTE: New samples may be tested in the same run with repeat samples.NOTE: In the occurrence of UNR or IND results impacting only one Master Mix, the test must be repeated with both Master Mixes clipped on the strip. In the event that the repeat yields a different reportable result for a given target, any positive result should be reported.Unresolved ResultUnresolved results may be obtained in the event that specimen-associated inhibition or a reagent failure prevents proper target or Sample Processing Control amplification. Sample(s) can be repeated from their corresponding Sample Buffer Tube(s) within the timeframes defined above. Vortex the sample(s) for 1 minute and restart following the BD MAX System Operation section.
10
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
Indeterminate ResultIndeterminate results may be obtained in the event that a System alert or a reagent failure occurs. Sample(s) can be repeated from their corresponding Sample Buffer Tube(s) within the timeframes defined above. Vortex the sample(s) for 1 minute and restart following the BD MAX System Operation section. For the interpretation of warning or error code messages, refer to the BD MAX System User’s Manual24 (Troubleshooting section).Incomplete ResultIncomplete results may be obtained in the event that Sample Preparation or the PCR failed to complete. Sample(s) can be repeated from their corresponding Sample Buffer Tube(s) within the timeframes defined above. Vortex the sample(s) for 1 minute and restart following BD MAX System Operation section. For the interpretation of warning or error code messages, refer to the BD MAX System User’s Manual24 (Troubleshooting section).External Control FailureExternal Controls should yield expected results when tested. If samples have to be repeated due to an incorrect External Control result, they should be repeated from their Sample Buffer Tubes along with freshly prepared External Controls within the timeframes defined above. Vortex the samples for 1 minute and restart following the BD MAX System Operation section.Interfering SubstancesTwenty-four(24)biologicalandchemicalsubstancesthatmaybepresentinvaginalspecimenswereevaluatedforpotentialinterference with the BD MAX Vaginal Panel. Exogenous (e.g. prescription and Over-the-Counter drugs, creams and/or gels) and endogenous (e.g. blood, hormones, mucus) were tested at the highest concentrations that may be found in vaginal specimens. KYJellyPersonalLubricantandWholeBloodwerefoundtointerfereatlevelsabove>12.5µL/mL. Zovirax Acyclovir 5% Cream and VCF Contraceptive Foam were found to interfere at levels above >3.1 µL/mL. MUKO Lubricating Jelly (Gel) was found to interfere above >3.3 µL/mL. Preparation H Hemorrhoidal Cream was found to interfere above >0.8 µL/mL. Interference with the following substanceswas observed at all tested levels: Conceptrol Vaginal Contraceptive Gel, Clotrimazole Vaginal Cream, Monistat 3 Cream, Vagisil Cream,Replens Vaginal Moisturizing Gel, Surgilube (Gel), Aquasonic Clear (Gel), McKesson Lubricating Jelly (Gel), Metronidazole, Leukocytes(Table 31). Substances that demonstrated interference may result in potential unresolved, indeterminate or false negative results.In addition, microorganisms that may be used in probiotics were evaluated for potential interference with the BD MAX Vaginal Panel. Fourteen (14) organisms were tested at high concentrations (>6.7 x 105 CFU/mL of Sample Buffer). Interference in the form of false negative BV results was observed in the presence of the following organisms: Lactobacillus amylovorus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus kefirgranum, and Lactobacillus helveticus.ResultsareshowninTable32.
Table 31: Exogenous and Endogenous Substances Tested for Interferencea
No Interference Observed Interference Observed
Substance Substance Level below Which No Interference Observed µL/mL)
Replens Vaginal Moisturizing GelMetronidazole 0.75% Gel
SurgilubeAquasonic Clear
McKesson Lubricating Jelly
Endo
geno
us Mucus (Bovine Cervical, 5% v/v) Whole Blood ≤12.5(equivalentto≤1.25%v/v)
Semen (5% v/v) Leukocytes Interference observed at each level evaluated
a In total, with the BD MAX Vaginal Panel in the presence of potentially interfering substances, 3,574 samples were tested for vaginosis targets and 4,159forvaginitistargets.Forvaginitistargets,ratesof10.12%INDand8.54%UNRresultswererecorded.ForBV,ratesof11.67%INDand2.57%UNR results were recorded.
11
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
Table 32: Microorganisms Tested for Interference
No Interference Observed Interference ObservedLactobacillus plantarum Lactobacillus casei Lactobacillus delbrueckii subsp. bulgaricus
Mixed Infection/Competitive InterferenceA mixed infection/competitive interference study was designed to evaluate the ability of the BD MAX Vaginal Panel to detect low positives in the presence of other targets at high concentrations. The following organisms and concentrations were evaluated.• Assay targets at high concentrations: L. crispatus (8.7 x 104 CFU/mL), G. vaginalis (5.0 x 106 CFU/mL), A. vaginae
(8.0 x 105 CFU/mL), Megasphaera-1(2.4x107 cp/mL), and T. vaginalis (3.3 x 105 to 1.0 x 106 cells/mL), C. albicans(1 x 106 CFU/mL), C. glabrata (1 x 106 CFU/mL)
• High concentrations of organisms of the vaginal flora: Dialister microaerophilus, Prevotella melaninogenica, Streptococcus mitis,Bifidobacterium breve and Mobiluncus curtisii (each at 1.0 x 106 CFU/mL).
• LowpositiveloadsofVaginitistargetsweretestedat<2xLoD.Samplescontaininglowpositiveloadsofbacterialvaginosismarkers were prepared with organism compositions sufficient to obtain 95% positive BV results.
A series of pools simulating mixed infections were tested: • BV analytes at high loads with low positive loads of:
○ T. vaginalis and C. albicans○ C. krusei and C. glabrata
• C. albicans high load with low positive loads of:○ C. krusei and C. glabrata○ T. vaginalis○ BV
• C. glabrata high load with low positive loads of:○ T. vaginalis○ BV
• C. krusei high load with low positive load of BV• T. vaginalis high load with low positive loads of:
○ BV○ C. albicans○ C. krusei○ C. glabrata
• Vaginal flora high load with low positive loads of:○ T. vaginalis and BV○ C. albicans and BV○ C. glabrata and BV○ C. krusei and BV
Samples containing T. vaginalis at low concentrations and low positive BV samples were successfully detected by the BD MAX Vaginal Panel when combined with high concentrations of other assay targets or with other organisms of the vaginal flora in the presence of simulated vaginal matrix. Competitive inhibition was observed for the following conditions:• For low positive samples containing Candida albicans,92%ofpositiveresultswereobtainedinpresenceofBVanalytesat
high loads.• For low positive samples containing Candida albicans, C. krusei or C. glabrata,BDMAXVaginalPanelgenerated42%,
61% and 33% of expected results respectively in presence of Trichomonas vaginalis at a load of 3.3 x 105 cells/mL.
12
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
LIMITATIONS OF THE PROCEDURE• The BD MAX Vaginal Panel is intended for use only with the BD MAX UVE Specimen Collection kit. • The BD MAX Vaginal Panel should only be used with the BD MAX System by trained personnel.• The BD MAX Vaginal Panel has not been validated for vaginal swab specimens collected by patients at home.• Collection and testing of patient-collected vaginal swab specimens with the BD MAX Vaginal Panel is not intended to replace
exam by a clinician. Vaginal infections may result from other causes or concurrent infections may occur. • Public health recommendations should be consulted regarding testing for additional sexually transmitted diseases for patients
with a positive result for Bacterial Vaginosis or T. vaginalis with the BD MAX Vaginal Panel.• Additional microorganisms not detected by the BD MAX Vaginal Panel such as Prevotella, Lachnospira, Sneathia, Mobiluncus,
Mycoplasma hominis, and Ureaplasma spp. have also been found in women with bacterial vaginosis, but are less associated with BV due to their relatively low prevalence, sensitivity and/or specificity.3,6,7
• Patients under 18 years old were not evaluated.• A Cgroup positive result can be due to one or multiple Candida species.• Reliable assay results are dependent on adequate specimen collection. Follow the procedures in this package insert and
the BD MAX UVE Specimen Collection Kit. Failure to follow specimen collection instructions may cause an increase in non-reportable results.
• Lubricants or other products containing substances such as carbomers, can increase the non-reportable rate obtained with BD MAX Vaginal Panel.
• Interference with the BD MAX Vaginal Panel was observed in the presence of the following substances: Conceptrol Vaginal Contraceptive Gel, Clotrimazole Vaginal Cream, Monistat 3 Cream, Vagisil Cream, Replens Vaginal Moisturizing Gel, Surgilube, McKesson Lubricating Jelly, Aquasonic Clear, Metronidazole, Leukocytes. The following substances were observed to interfere at levels above the stated concentrations: Preparation H Hemorrhoidal Cream (>0.8 µL/mL), Zovirax Acyclovir 5% Cream (>3.1µL/mL),VCFContraceptiveFoam(>3.1µL/mL),KYJellyPersonalLubricant(>12.5µL/mL),MUKOLubricatingJelly(>3.3µL/mL),WholeBlood(>12.5µL/mLor>1.25%v/v).
• Interference with the BD MAX Vaginal Panel was observed in the presence of the following microorganisms: Lactobacillus amylovorus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus kefirgranum, and Lactobacillus helveticus, that may all be used in probiotics. Interference with Lactobacillus kefiranofaciens has not been determined.
• Cross-reactivity with the BD MAX Vaginal Panel may be observed with the following organisms: Lactobacillus acidophilus, detected in human stool; Trichomonas tenax, a commensal of the oral cavity.
• The following organisms were observed to cross-react above the stated concentrations: Olsenella uli (>6.6 x 104 CFU/mL) and Atopobium rimae (>4.4 x 104 CFU/mL), both isolated from oral cavity; Lactobacillus delbrueckii subsp. lactis (>3.9 x 103 CFU/mL), detected in human stool; Pichia fermentans (>6.0 x 103 CFU/mL), from coffee beans and fruit.
• All strains tested in the Inclusivity study were detected. However, Three (3) out of 10 tested Gardnerella vaginalis strains and 1 out of 5 Lactobacillus crispatus strains were detected only at high concentrations (9x LoD and 5x LoD respectively).
• The effects of other potential variables such as vaginal discharge, use of tampons and specimen collection variables have not been determined.
• As with many diagnostic tests, results from the BD MAX Vaginal Panel should be interpreted in conjunction with other laboratory and clinical data available to the physician.
• Candida species can be present as commensal organisms in a significant percentage of women and BD MAX Vaginal Panel results should be considered in conjunction with other clinical and patient information to determine the disease status.
• Bacterial vaginosis marker combinations that generate positive results for bacterial vaginosis can be commensal in a significant percentage of women and therefore positive results for bacterial vaginosis should be considered in conjunction with other clinical and patient information to determine the disease status.
• Erroneous test results may occur from improper specimen collection, handling or storage, technical error, sample mix-up or because the number of organisms in the sample is below the analytical sensitivity of the test.
• If the BD MAX Vaginal Panel result is IND, INC, or UNR (for one or more targets) then the test should be repeated.• Good laboratory technique is essential for the proper performance of this assay. Due to the high analytical sensitivity of this test,
extreme care should be taken to preserve the purity of all materials and reagents.• A positive test result does not necessarily indicate the presence of viable organisms. A positive result is indicative of the
presence of target DNA. • The BD MAX Vaginal Panel cannot be used to assess therapeutic success or failure since target nucleic acids may persist
following antimicrobial therapy.• As with all PCR-based tests, extremely low levels of target below the LoD of the assay may be detected, but results may not
be reproducible.
13
BD MAX™ Vaginal PanelCLSI Laboratory Procedure
• False negative results may occur due to loss of nucleic acid from inadequate collection, transport or storage of specimens,or due to inadequate cell lysis. The Sample Processing Control has been added to the test to aid in the identification ofspecimens that contain inhibitors to PCR amplification. The Sample Processing Control does not indicate if nucleic acid hasbeen lost due to inadequate collection, transport or storage of specimens, or if cells have been adequately lysed.
• At very low load, false negative Candida glabrata results may occur for specimens containing more than one analyte.• At very low load, false negative Candida spp. results may occur for specimens containing more than one BV analyte at
high loads.• At very low load, false negative Candida spp., C. krusei or C. glabrata results may occur for specimens containing high load of
T. vaginalis. This test is a qualitative test and does not provide quantitative values nor indicate the quantity of organisms present.• Mutations or polymorphisms in primer- or probe-binding regions may affect detection resulting in a false negative result with the
BD MAX Vaginal Panel.• This test is a qualitative test and does not provide quantitative values nor indicate the quantity of organisms present.
bacterial vaginosis.” Clin Infect Dis 47(1): 33–43.5. McCormack WM. “Vulvovaginitis and cervicitis”. In: Mandell GL, Bennett JE, Dolin R, (eds). Principles and Practice of Infectious
molecular diagnosis of bacterial vaginosis is possible?” PLoS One 8(4): e60670.7. SrinivasanS,HoffmanNG,etal.(2012).“Bacterialcommunitiesinwomenwithbacterialvaginosis:highresolutionphylogenetic
analyses reveal relationships of microbiota to clinical criteria.” PLoS One 7(6): e37818.8. CentersforDiseaseControlandPrevention.SexuallyTransmittedDiseasesTreatmentGuidelines,2015.MMWRRecommRep
multiplexPCR.”InfectDisObstetGynecol2012:872169,doi:10.1155/2012/872169.16. Fidel Jr. PL, Vazquez JA, et al. (1999). “Candida glabrata: review of epidemiology, pathogenesis, and clinical disease with
standard.” Obstet Gynecol. 113(1):89–95.22. Clinical and Laboratory Standards Institute. Protection of laboratory workers from occupationally acquired infections; Approved
Guideline.DocumentM29(Refertothelatestedition).23. Centers for Disease Control and Prevention, and National Institutes of Health. Biosafety in microbiological and biomedical
24. BDMAXSystemUser’sManual(Refertothelatestversion)BDLifeSciences,Sparks,MD21152USA.25. Clinical and Laboratory Standards Institute. Molecular Diagnostic Methods for Infectious Diseases; Approved Guideline,
Document MM3 (Refer to the latest edition).26. ClinicalandLaboratoryStandardsInstitute.2006.ApprovedGuideline,DocumentC24.StatisticalQualityControlfor
Quantitative Measurements: Principles and Definitions, 3rd ed., CLSI. Wayne PA (Refer to latest edition).27. Clinical and Laboratory Standards Institute. User Protocol for Evaluation of Qualitative Test Performance; Approved Guideline,
![l>lf·· E ·B; -I,:,C-·-1·1V · cat. no.i bd lj.657 bd lj.6]5 bd 4630 bd 4·627 bd 4628 bd 4886 bd 4546 bd 4·545 bd 4544 bd 4542 bd lj,588 bd lj.593 bd 0102 bd 4636 bd 4632 bd](https://static.documents.pub/doc/80x56/5f7c69bb7d840d18665ab1e6/llf-e-b-ic-11v-cat-noi-bd-lj657-bd-lj65-bd-4630-bd-4627-bd-4628-bd.jpg)
