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
 14 human species; three pathogenic
 M. hominis
 M. genitalium
 M. pneumoniae
Ureaplasma
Microscopic view of Mycoplamsas
 2 human species; both pathogenic
 U. urealyticum
 U. parvum
(Taylor-Robinson et al., An International Journal of Obstetrics and Gynecology,
2010)
LABORATORY CHARACTERISTICS
 Facultative anaerobes
 Pleomorphic
 Limited genome
 Unable to gram stain
 Culture?
(Larsen et al., Infectious Diseases in Obstetrics and Gynecology, 2010)
VIRULENCE
 Normal Flora/Non-pathogenic
colonizers?
 Pathogenic?
 Opportunistic pathogen






Location of colonization
Host immune response
Conditions of pregnancy
Co-infections
Genetic factors
Environmental factors
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.
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
( K a t a o ka , J o u r n a l o f C l i n i c a l M i c r o b i o l o gy, 2 0 0 6 ) , ( Ta yl o r - R o b i n s o n , An I n t e r n a t i o n a l J o u r n a l o f O b s t e t r i c s a n d Gyn e c o l o g y,
2010)
MYCOPLASMA HOMINIS
 Strongly associated with:
 Chorioamnionitis
 Pelvic inflammatory disease
 Bacterial vaginosis
 Pregnancy
Mycoplasma hominis on agar plate




Lower gestational age at delivery
Lower birth weight
Increased neonatal morbidity & mortality
Increase risk for miscarriage at 14 weeks
 Infant
 Pneumonia
MYCOPLASMA GENITALIUM
 Causative agent of urethritis
 Associated with cervicitis, PID, and
endometritis in women
 Pregnancy- Unknown
Electron micrograph of M.
genitalium
( Ta yl o r - R o b i n s o n , An I n t e r n a t i o n a l J o u r n a l o f O b s t e t r i c s a n d Gyn e c o l o g y, 2 0 1 0 )
UREAPLASMA SPP.
In 2002, U. urealyticum & U. parvum
distinguished as separate species
Therefore, studies pre-2002 confounded
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
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:
Electron micrograph of U. parvum
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

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PROM
Preterm labor
Chorioamnionitis (in mother)
Early onset sepsis & BPD (bronchopulmonary
dysplasia) in baby
( La r s e n e t a l . , I n fe c t i o u s D i s e a s e s i n O b s t e t r i c s a n d Gyn e c o l o g y, 2 0 1 0 )
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
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
RESULTS
Bacterial characteristics and cause of early delivery for the preterm birth group.
Subject
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
a
Gestational wk at delivery
6
9
18
26
30
31
34
35
35
35
35
35
36
36
36
36
36
36
36
36
36
36
36
Presence (+/-) of:
U. parvum
U. urealyticum
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
-
Cause of
preterm deliverya
Miscarriage
Miscarriage
Miscarriage
PROM
Preeclampsia
PROM
FSUA
Preeclampsia
Preeclampsia
FSUA
FSUA
FSUA
FSUA
FSUA
FSUA
FSUA
FSUA
FSUA
FGR
Preeclampsia
Preeclampsia
FSUA
FSUA
PROM, premature rupture of the membranes; FSUA, failure to suppress uterine activity; FGR, fetal growth restriction.
RESULTS
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
Preterm birth (20)
2 (10)
0.999
Preterm labor (4)
0
0.999
3 (100)
15 (75)
4 (100)
•27 abnormal pregnancy outcomes -25 associated with U. parvum
•U. parvum strongly associated with their occurrence (p=0.003)
0.245
0.057
0.122
CURRENT STUDY
 Previous study
 Small population sample
 Little diversity & limited risk factors
Parameters:
 Project collaboration with WiNHR
( Wi s cons in N et w o r k f o r H eal t h care Res ear ch)
 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
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
HYBRIDIZATION ASSAY
Biotinylated
amplification product
Aminated probe
(4 species specific probes used)
HYBRIDIZATION ASSAY
Biotinylated
amplification product
Aminated probe
(4 species specific probes used)
HYBRIDIZATION ASSAY
E
Strepavidin-HPO conjugate
Biotinylated
amplification product
Aminated probe
(4 species specific probes used)
HYBRIDIZATION ASSAY
Substrate
E(S)
Strepavidin-HPO conjugate
Biotinylated
amplification product
Aminated probe
(4 species specific probes used)
STUDY PROGRESS
 PCR hybridization assay completed on samples
 Correlations between species and adverse pregnancy
outcomes
ACKNOWLEDGEMENTS
 Thanks to:
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Microbiology Research Laboratory
Gundersen Lutheran Medical Foundation
Dr. Steve Callister
Dean Jobe