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ASSOCIATION BETWEEN MYCOPLASMA INFECTION AND COMPLICATIONS DURING PREGNANCY

ASSOCIATION BETWEEN MYCOPLASMA INFECTION AND COMPLICATIONS DURING PREGNANCY. Steven Lovrich , Gundersen Lutheran Medical Foundation. MYCOPLASMA. Mollicutes : “soft skin” Intracellular parasite Lack cell wall Trilayered external membranes 2 genera: Mycoplasma

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ASSOCIATION BETWEEN MYCOPLASMA INFECTION AND COMPLICATIONS DURING PREGNANCY

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  1. ASSOCIATION BETWEEN MYCOPLASMA INFECTION AND COMPLICATIONS DURING PREGNANCY Steven Lovrich, Gundersen Lutheran Medical Foundation

  2. MYCOPLASMA • 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) Microscopic view of Mycoplamsas

  3. LABORATORY CHARACTERISTICS • Facultative anaerobes • Pleomorphic • Limited genome • Unable to gram stain • Culture? (Larsen et al., Infectious Diseases in Obstetrics and Gynecology, 2010)

  4. VIRULENCE • Normal Flora/Non-pathogenic colonizers? • Pathogenic? • Opportunistic pathogen • Location of colonization • Host immune response • Conditions of pregnancy • Co-infections • Genetic factors • Environmental factors

  5. PATHOGENICITY DURING PREGNANCY • 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.

  6. PRE-TERM BIRTH 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)

  7. MYCOPLASMA HOMINIS • 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 on agar plate

  8. MYCOPLASMA GENITALIUM • Causative agent of urethritis • Associated with cervicitis, PID, and endometritis in women • Pregnancy- Unknown (Taylor-Robinson, An International Journal of Obstetrics and Gynecology, 2010) Electron micrograph of M. genitalium

  9. UREAPLASMA SPP. • In 2002, U. urealyticum & U. parvum distinguished as separate species • Therefore, studies pre-2002 confounded

  10. UREAPLASMA UREALYTICUM • 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 Electron micrograph of U. urealyticum

  11. UREAPLASMA PARVUM • 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) Electron micrograph of U. parvum

  12. ASSOCIATION OF UREAPLASMA WITH PRETERM BIRTH 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

  13. SUMMARY OF RESULTS • 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

  14. 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.

  15. Results • 27 abnormal pregnancy outcomes -25 associated with U. parvum • U. parvum strongly associated with their occurrence (p=0.003)

  16. CURRENT STUDY • 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

  17. EXPERIMENTAL DESIGN 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

  18. Hybridization assay Biotinylated amplification product Aminated probe (4 species specific probes used)

  19. Hybridization assay Biotinylated amplification product Aminated probe (4 species specific probes used)

  20. Hybridization assay Strepavidin-HPO conjugate E Biotinylated amplification product Aminated probe (4 species specific probes used)

  21. Hybridization assay Substrate Strepavidin-HPO conjugate E(S) Biotinylated amplification product Aminated probe (4 species specific probes used)

  22. STUDY PROGRESS • PCR hybridization assay completed on samples • Correlations between species and adverse pregnancy outcomes

  23. Acknowledgements • Thanks to: • Microbiology Research Laboratory • Gundersen Lutheran Medical Foundation • Dr. Steve Callister • Dean Jobe

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