Post on 12-Jan-2016
transcript
Steven Lovrich, Gundersen Lutheran Medical Foundation
ASSOCIATION BETWEEN MYCOPLASMA INFECTION
AND COMPLICATIONS DURING PREGNANCY
Mollicutes: “soft skin” Intracellular parasite Lack cell wall Trilayered external membranes
2 genera:Mycoplasma
14 human species; three pathogenic M. hominis M. genitalium M. pneumoniae
Ureaplasma 2 human species; both pathogenic
U. urealyticum U. parvum
(Taylor-Robinson et al., An International Journal of Obstetrics and Gynecology, 2010)
MYCOPLASMA
Microscopic view of Mycoplamsas
Facultative anaerobes
Pleomorphic
Limited genome
Unable to gram stain
Culture?
(Larsen et al., Infectious Diseases in Obstetrics and Gynecology, 2010)
LABORATORY CHARACTERISTICS
Normal Flora/Non-pathogenic colonizers?
Pathogenic?
Opportunistic pathogen Location of colonization Host immune response Conditions of pregnancy Co-infections Genetic factors Environmental factors
VIRULENCE
Adherence to host cell by mycoplasmal adhesion proteins/lipoproteins
Stimulate secretion of pro-inflammatory cytokines (tumor necrosis factor-, interleukin, & interferon-γ)
Stimulate release of prostaglandins which leads to protease production
Protease can cause adverse pregnancy outcomes miscarriage, pre-term labor, bacterial vaginosis, chorioamnionitis,
spontaneous abortion, perinatal morbidity & mortality, PROM, etc.
PATHOGENICITY DURING PREGNANCY
Problems:Genital tract infections associated with approximately 50% of
preterm deliveries
13% of pregnancies in the U.S. result in preterm delivery or low infant birth weight
60% of mortality among infants (with no anatomic/chromosomal defects) is low birth weight
(Kataoka, Journal of Clinical Microbiology, 2006), (Taylor-Robinson, An International Journal of Obstetrics and Gynecology, 2010)
PRE-TERM BIRTH
Strongly associated with: Chorioamnionitis Pelvic inflammatory disease Bacterial vaginosis
Pregnancy Lower gestational age at delivery Lower birth weight Increased neonatal morbidity & mortality Increase risk for miscarriage at 14 weeks
Infant Pneumonia
MYCOPLASMA HOMINIS
Mycoplasma hominis on agar plate
Causative agent of urethritis
Associated with cervicitis, PID, and endometritis in women
Pregnancy- Unknown
(Taylor-Robinson, An International Journal of Obstetrics and Gynecology, 2010)
MYCOPLASMA GENITALIUM
Electron micrograph of M. genitalium
In 2002, U. urealyticum & U. parvum distinguished as separate species
Therefore, studies pre-2002 confounded
UREAPLASMA SPP.
Colonization of placenta= increases risk for fetal & maternal inflammation Increase risk of preterm labor Increase risk for miscarriage (@14 weeks)
Vertically transmitted to fetus potentially causing: Bacteremia Pneumonia Chronic lung disease Nervous system infections
UREAPLASMA UREALYTICUM
Electron micrograph of U. urealyticum
Vertically transmitted to fetus in utero or during delivery Bacteremia, pneumonia, chronic lung disease, & nervous
system infections
More prevalent in amniotic fluid of preterm pregnancies than U. urealyticum
If colonization occurs can cause: PROM Preterm labor Chorioamnionitis (in mother) Early onset sepsis & BPD (bronchopulmonary dysplasia) in
baby
(Larsen et al., Infectious Diseases in Obstetrics and Gynecology, 2010)
UREAPLASMA PARVUM
Electron micrograph of U. parvum
Objectives:
1. Determine if colonization of either Ureaplasma species had association with miscarriage or preterm labor
2. To perfect detection methods and discrimination of species
Methods: Tested 239 pregnant women (PCR) from the
La Crosse area for colonization with Ureaplasma
urealyticum & parvum during early prenatal period
ASSOCIATION OF UREAPLASMA WITH PRETERM BIRTH
239 patient samples at start
192 follow ups at Gundersen Lutheran 47 lost
27 adverse events 23 preterm birth (≤36 weeks) or miscarriage 4 preterm labor (stopped)
Significance of Colonization P-value ≤ 0.05
SUMMARY OF RESULTS
RESULTS
Bacterial characteristics and cause of early delivery for the preterm birth group. Presence (+/-) of: Cause of
Subject Gestational wk at delivery U. parvum U. urealyticum preterm deliverya
1 6 + - Miscarriage 2 9 + - Miscarriage 3 18 + - Miscarriage 4 26 + - PROM 5 30 + - Preeclampsia 6 31 - - PROM 7 34 + - FSUA 8 35 + + Preeclampsia 9 35 + - Preeclampsia 10 35 + - FSUA 11 35 + - FSUA 12 35 + - FSUA 13 36 + - FSUA 14 36 + - FSUA 15 36 - + FSUA 16 36 - - FSUA 17 36 - - FSUA 18 36 + - FSUA 19 36 + - FGR 20 36 + - Preeclampsia 21 36 + - Preeclampsia 22 36 - - FSUA 23 36 + - FSUA
a PROM, premature rupture of the membranes; FSUA, failure to suppress uterine activity; FGR, fetal growth restriction.
Table 2. Association between urogenital infection with ureaplasmas and miscarriage, preterm
labor controlled by treatment, and/or preterm birth.
Pregnancy outcome (no.) No. (%) U. urealyticum p value No. (%) U. parvum p value Normal (165) 16 (10) NA 84 (51) NA Abnormal (27) 2 (7) 0.999 22 (81) 0.003
Miscarriage (3) 0 0.999 3 (100) 0.245 Preterm birth (20) 2 (10) 0.999 15 (75) 0.057 Preterm labor (4) 0 0.999 4 (100) 0.122
RESULTS
•27 abnormal pregnancy outcomes -25 associated with U. parvum•U. parvum strongly associated with their occurrence (p=0.003)
Previous study Small population sample Little diversity & limited risk factors
Parameters:Project collaboration with WiNHR
(Wisconsin Network for Healthcare Research)
4 different hospital sites: 200 samples per site Aurora Health, Gundersen Lutheran, UW-Hospital (Madison), Marshfield Cinic
Test for 4 Mycoplasma species
*Objective: To determine if any of the 4 Mycoplasma species correlate with pregnancy abnormalities or adverse outcomes Examine multiple risk factors
CURRENT STUDY
Normal healthy pregnant women targeted in 4 sites across Wisconsin
Swabs of urogenital tract (~12 weeks)
Samples blinded
DNA extracted and forwarded to Gundersen Lutheran
Polymerase Chain Reaction (PCR)
Hybridization assay
EXPERIMENTAL DESIGN
HYBRIDIZATION ASSAY
Aminated probe(4 species specific probes used)
Biotinylatedamplification product
HYBRIDIZATION ASSAY
Aminated probe(4 species specific probes used)
Biotinylatedamplification product
HYBRIDIZATION ASSAY
E
Aminated probe(4 species specific probes used)
Biotinylatedamplification product
Strepavidin-HPO conjugate
HYBRIDIZATION ASSAY
E(S)
Aminated probe(4 species specific probes used)
Biotinylatedamplification product
Strepavidin-HPO conjugate
Substrate
PCR hybridization assay completed on samples
Correlations between species and adverse pregnancy outcomes
STUDY PROGRESS
Thanks to: Microbiology Research Laboratory Gundersen Lutheran Medical Foundation Dr. Steve Callister Dean Jobe
ACKNOWLEDGEMENTS