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Microbial prevalence, diversity and abundance in amniotic fluid during preterm labor: a molecular and culture-based investigation

Microbial prevalence, diversity and abundance in amniotic fluid during preterm labor: a molecular and culture-based investigation

Authors :
Roberto Romero
Elisabeth M. Bik
Chong Jai Kim
Sam Edwin
Francesca Gotsch
Juan Pedro Kusanovic
Harold P. Amogan
Daniel B. DiGiulio
Offer Erez
David A. Relman
Source :
PLoS ONE, Vol 3, Iss 8, p e3056 (2008), PLoS ONE
Publication Year :
2008
Publisher :
Public Library of Science (PLoS), 2008.

Abstract

Background Preterm delivery causes substantial neonatal mortality and morbidity. Unrecognized intra-amniotic infections caused by cultivation-resistant microbes may play a role. Molecular methods can detect, characterize and quantify microbes independently of traditional culture techniques. However, molecular studies that define the diversity and abundance of microbes invading the amniotic cavity, and evaluate their clinical significance within a causal framework, are lacking. Methods and Findings In parallel with culture, we used broad-range end-point and real-time PCR assays to amplify, identify and quantify ribosomal DNA (rDNA) of bacteria, fungi and archaea from amniotic fluid of 166 women in preterm labor with intact membranes. We sequenced up to 24 rRNA clones per positive specimen and assigned taxonomic designations to approximately the species level. Microbial prevalence, diversity and abundance were correlated with host inflammation and with gestational and neonatal outcomes. Study subjects who delivered at term served as controls. The combined use of molecular and culture methods revealed a greater prevalence (15% of subjects) and diversity (18 taxa) of microbes in amniotic fluid than did culture alone (9.6% of subjects; 11 taxa). The taxa detected only by PCR included a related group of fastidious bacteria, comprised of Sneathia sanguinegens, Leptotrichia amnionii and an unassigned, uncultivated, and previously-uncharacterized bacterium; one or more members of this group were detected in 25% of positive specimens. A positive PCR was associated with histologic chorioamnionitis (adjusted odds ratio [OR] 20; 95% CI, 2.4 to 172), and funisitis (adjusted OR 18; 95% CI, 3.1 to 99). The positive predictive value of PCR for preterm delivery was 100 percent. A temporal association between a positive PCR and delivery was supported by a shortened amniocentesis-to-delivery interval (adjusted hazard ratio 4.6; 95% CI, 2.2 to 9.5). A dose-response association was demonstrated between bacterial rDNA abundance and gestational age at delivery (r2 = 0.42; P

Details

Language :
English
ISSN :
19326203
Volume :
3
Issue :
8
Database :
OpenAIRE
Journal :
PLoS ONE
Accession number :
edsair.doi.dedup.....490013127db4a040637aa1821cfd3ded