Date post: | 11-Nov-2014 |
Category: |
Health & Medicine |
Upload: | umang31 |
View: | 400 times |
Download: | 0 times |
UTI-lab diagnosis MGMC&H,Jaipur 1
Laboratory Diagnosis
URINARY TRACT INFECTION
UTI-lab diagnosis MGMC&H,Jaipur 2
Introduction• It is defined as a disease caused by microbial invasion of the genitourinary tract that extends from the renal cortex of the kidney to the urethral meatus.
•Route of infection to reach the urinary tract can be either- Ascending route Haematogenous route
UTI-lab diagnosis MGMC&H,Jaipur 3
Causative organisms
• E.coli• Klebsiella• Pseudomonas• Proteus
Gram –ve Bacilli
• Staphylococcus• Enterococci
Gram +ve Bacilli
• M.tuberculosis• Citrobacter• Salmonella
Miscellaneous
• Candida albicans
Fungus
E.coli
c.albicans
Citrobacter
Staphylococcus
UTI-lab diagnosis MGMC&H,Jaipur 4
UTI causing organisms
E.coli (63%)Klebssiella(15%)Pseudomonas(5%)Proteus(5%)Other Gram -ve bac-teriaStaphylococcusEnterococcus
UTI-lab diagnosis MGMC&H,Jaipur 5
Sterile pyuria•Sterile pyuria is urine which contains white blood cells while appearing sterile by standard culturing techniques.
• Sterile pyuria is listed as a side effect from some medications such as paracetamol (acetaminophen).
UTI-lab diagnosis MGMC&H,Jaipur 6
Community acquired UTI
•An infection contracted outside of a health care setting or an infection present on admission.•E.Coli is the most common cause followed by stapylococcus saprophyticus
s.saprophyticus
UTI-lab diagnosis MGMC&H,Jaipur 7
Hospital acquired UTI•Acquired by a patient during a hospital visit.•Most common organism causing it- Klebsiella
UTI-lab diagnosis MGMC&H,Jaipur 8
Patients with urinary catheters have a higher chance of getting UTI’s. This is because germs can travel along the tubing of the catheter and cause infection in the urinary system. This is then referred to as a “catheter associated” UTI.
Complications –•Cystitis• pyelonephritis• bacteremia• prostatitis•Rarely, endocarditis, septic arthritis, and meningitis
Catheter acquiredUTI
UTI-lab diagnosis MGMC&H,Jaipur 9
Poor aseptic insertion of catheter: periurethral flora inserted into bladder.
Migration of bacteria along outer surface of catheter.
Open drainage.
Breaks in the closed drainage system.
Causes
UTI-lab diagnosis MGMC&H,Jaipur 10
SPECIMEN COLLECTION
TRANSPORT and STORAGE
LABORATORY METHODS
LABDIAGNOSIS
1. 2.
3.
UTI-lab diagnosis MGMC&H,Jaipur 11
SPECIMEN COLLECTION
UTI-lab diagnosis MGMC&H,Jaipur 12
MID STREAM SPECIMEN(MSU)•Collected PRIOR to antibiotics.
•WASH hands properly.•Specimen is collected in a STERILE container
BEFORE ANTIBIOTICS
STERILE CONTAINER
UTI-lab diagnosis MGMC&H,Jaipur 13
Method-
First morning specimen should be taken.
FEMALES- MALES-
Clean the area with soap and water , hold the labia apart and discard few ml of urine and then collect the midstream sample directly into the sterile container.
Clean the glans with soap and water, retract the foreskin, discard few ml of urine and the collect the mid stream sample directly into the sterile container .
•Discard the rest of the urine.•The first portion of urine adequately flushes out the normal urethral flora.
UTI-lab diagnosis MGMC&H,Jaipur 14
CATHETER SPECIMEN
•Urine should be collected directly from the catheter and not from the collection bag.
•The catheter should not touch the container.
•Catheterization to obtain urine is not done because of risk of introducing infection though it yields excellent results.
URINE SPECIMEN FROM INFANTS
A clean catch specimen after cleansing of genitalia. OR Supra pubic aspiration.
UTI-lab diagnosis MGMC&H,Jaipur 15
TRANSPORTand
STORAGE
UTI-lab diagnosis MGMC&H,Jaipur 16
THE SPECIMEN SHOULD BE TRANSPORTED IMMEDIATELY.
•Urine is a good medium for the growth of microorganisms, hence delay in processing will hamper the results.
•If delay is unavoidable, the specimen can be refrigerated for up to four hours at 4˚c.
UTI-lab diagnosis MGMC&H,Jaipur 17
Urine transport tubes have shown to preserve bacteria without refrigeration.It consistes of-
-Boric acid -Glycerol
-Sodium formate
UTI-lab diagnosis MGMC&H,Jaipur 18
LABORATORY
METHODSMICROSCOPYCULTUREIDENTIFICATIONOTHER METHODSANTIBIOTIC SENSTIVITY TEST
UTI-lab diagnosis MGMC&H,Jaipur 19
MICROSCOPYUrine is centrifuged and deposit is examined under the microscope.
Following can be seen-•Pus cell( 0-5/hpf is the normal value for pus cells in the human body).
•RBC
•Epithelial cells
•Bacteria
Bacteria
Pus cell
UTI-lab diagnosis MGMC&H,Jaipur 20
CULTUREMost lab use a semi quantitative method (STANDARD LOOP TECHNIQUE) for culture.
This technique involves a standard calibrated loop which transfers a fixed and small volume of urine.
STANDARD CALIBRATED LOOP
UTI-lab diagnosis MGMC&H,Jaipur 21
Fixed and small amount of uncentrifuged urine is transferred to BLOOD and MacCONKEY’S AGAR.
Next day, the number of colonies grown is counted and total count per ml is calculated.
Standard loop technique
•BLOOD AGAR- gives a quantitative measurement of bacteriuria.•Mac CONKEY’S AGAR- gives presumptive diagnosis of the bacteria.
UTI-lab diagnosis MGMC&H,Jaipur 22
INTERPRETATION OF RESULTS
•Bacteriological diagnosis of UTI is carried out by ‘SIGNIFICANT BACTERIURIA’ using quantitative cultures developed by KASS(1956).
•This is based on the fact that normal urine is sterile but during voiding may become contaminated with genital commensals.
SIGNIFICANT BACTERIURIA
DOUBTFUL SIGNIFICANCE
NO SIGNIFICANT GROWTH
•When bacterial count is more than 105/ml of a single species.
•It indicates ACTIVE UTI.
•Between 104 to 105 bacteria per ml.
•Specimen should be repeated for culture.
• <103 bacteria per ml are regarded as contaminated.
• Contamination is also considered when 3 or more bacteria are isolated.
UTI-lab diagnosis MGMC&H,Jaipur 23
IDENTIFICATION
The organisms are identified by-
colony characters gram staining motility biochemical test slide agglutination test serological test
UTI-lab diagnosis MGMC&H,Jaipur 24
SIZE 1-2 mm
COLONY CHARACTER Blood agar
Mac Conkey agar
Grey-white, moist, smooth, opaque, non mucoid Pink colony( due to lactose fermentation)
GRAM STAINING Gram –ve bacteria
BIOCHEMICAL TEST Indole Methyl red Voges-Proskauer(VP) Citrate
++--
MOTILITY Motile
E.coli
UTI-lab diagnosis MGMC&H,Jaipur 25
BLOOD AGAR MacCONKEY’S AGAR
GRAM STAINING
UTI-lab diagnosis MGMC&H,Jaipur 26
SIZE 2-4 mm
COLONY CHARACTER Blood agar
Mac Conkey agar
Grey-white, moist, smooth, opaque, mucoid Pink colony( due to lactose fermentation)
GRAM STAINING Gram –ve bacteria
BIOCHEMICAL TEST Indole Methyl red Voges-Proskauer(VP) Citrate
--++
MOTILITY Non-Motile
Klebsiella
UTI-lab diagnosis MGMC&H,Jaipur 27
BLOOD AGAR
GRAM STAINING
MacCONKEY’S AGAR
UTI-lab diagnosis MGMC&H,Jaipur 28
ProteusCOLONY CHARACTER Blood agar Nutrient agar Mac Conkey agar
Exhibits SWARMING(fishy odour)Exhibits SWARMINGSmooth pale colonies
GRAM STAINING Gram –ve bacteria
BIOCHEMICAL TEST Indole TSI PPA Urease Oxidase
-(P.mirabilis)+(P.vulgaris)H2S production++-
MOTILITY Motile
UTI-lab diagnosis MGMC&H,Jaipur 29
BLOOD AGAR
GRAM STAINING
NUTRIENT AGAR
UTI-lab diagnosis MGMC&H,Jaipur 30
PseudomonasSIZE 2-4mm
COLONY CHARACTER Blood agar Nutrient agar Mac Conkey agar
Smooth, large, convex, translucent(sweetish aromatic odour)Same as on blood agarSmooth pale colonies
GRAM STAINING Gram –ve bacteria
BIOCHEMICAL TEST Indole TSI PPA Urease OxidaseCitrate
-K/ no change (without H2S production)-+/-+-
MOTILITY Motile
UTI-lab diagnosis MGMC&H,Jaipur 31
BLOOD AGAR
GRAM STAINING
NUTRIENT AGAR
UTI-lab diagnosis MGMC&H,Jaipur 32
OTHER METHODS-
Dip slides
Pour plate method
Triphenyltetrazolium
chloride (TTC) test
Griess nitrite test
Glucose test paper
Polymorphonuclear
neutrophils (PMNs)
Catalase test
Leucocyte esterase
Detection of lipopolysaccha
ride (endotoxin)
UTI-lab diagnosis MGMC&H,Jaipur 33
DIP SLIDES
In this test commercially available plastic slides coated with CLED agar (cysteine lactose electrolyte deficient) on one side and MacConkey’s agar on the other is dipped in urine then placed in a sterile container is incubated at 37˚c.
UTI-lab diagnosis MGMC&H,Jaipur 34
POUR PLATE METHOD•It is a quantitative method •But, is cumbersome for routine diagnostic work
UTI-lab diagnosis MGMC&H,Jaipur 35
GRIESS NITRITE TEST
•It is based on nitrate reducing enzymes produced by bacteria present in urine.
•Gram -ve bacteria- NITRATES NITRITES
•The presence of nitrite detected by the test indicates UTI.
•Normal urine does not contain nitrite.
UTI-lab diagnosis MGMC&H,Jaipur 36
TRIPHENYLTETRAZOLIUM CHLORIDE TEST (TTC TEST)
TTC is reduced by several frequently encountered urinary tract pathogens and produce a pink red precipitate.
UTI-lab diagnosis MGMC&H,Jaipur 37
CATALASE TEST•Certain bacteria have an enzyme catalase which acts on hydrogen peroxide to release oxygen.
H2O2 H2O + O
[Nascent
oxygen]
•The presence of catalase is evident by formation of bubbles.•It indicates bacteriuria. (test present only in catalase +ve organisms.)
catalase
UTI-lab diagnosis MGMC&H,Jaipur 38
GLUCOSE TEST PAPER
It is based on utilisation of the minute amount of glucose present in normal urine, by bacteria causing the infection.
Hence, it indicates bacteriuria.
UTI-lab diagnosis MGMC&H,Jaipur 39
POLYMORPHONUCLEAR NEUTOPHILS(PMNs)
•PMNs are counted in uncentrifuged urine specimen with the help of haemocytometer.
8 PMN/mm3 is indicative of infection.
UTI-lab diagnosis MGMC&H,Jaipur 40
LEUCOCYTE ESTERASEPresence of the enzyme in urine indicates infection.
DETECTION OF LIPOPOLYSACCHARIDE
This test helps in the detection of gram –ve bacteria in urine.
UTI-lab diagnosis MGMC&H,Jaipur 41
ANTIBIOTIC SENSITIVITY TEST
•Esch. coli and other common urinary pathogens develop multiple drug resistance and of the transferable variety.
•Antibiotic sensitivity is necessary to administer proper antibiotic.
•Done by STOKES DISC DIFFUSION method.
UTI-lab diagnosis MGMC&H,Jaipur 42
-Umang Juneja