Prevalence and Diversity of Microbes in the Amniotic Fluid, the Fetal Inflammatory Response, and Pregnancy Outcome in Women with Preterm Pre-Labor Rupture of Membranes

Authors

  • Daniel B. DiGiulio,

    1. Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA
    2. Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA
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  • Roberto Romero,

    1. Perinatology Research Branch, NICHD, NIH, Bethesda, MD, and Detroit MI, USA
    2. Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, MI, USA
    3. Department of Obstetrics and Gynecology, Wayne State University School of Medicine, Hutzel Women’s Hospital, Detroit Medical Center, Detroit, MI, USA
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  • Juan Pedro Kusanovic,

    1. Perinatology Research Branch, NICHD, NIH, Bethesda, MD, and Detroit MI, USA
    2. Department of Obstetrics and Gynecology, Wayne State University School of Medicine, Hutzel Women’s Hospital, Detroit Medical Center, Detroit, MI, USA
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  • Ricardo Gómez,

    1. Department of Obstetrics and Gynecology, CEDIP (Center for Perinatal Diagnosis and Research), Sotero del Rio Hospital, Santiago, Chile
    2. Department of Obstetrics and Gynecology, P. Universidad Católica de Chile, Santiago, Chile
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  • Chong Jai Kim,

    1. Perinatology Research Branch, NICHD, NIH, Bethesda, MD, and Detroit MI, USA
    2. Department of Pathology, Wayne State University School of Medicine, Detroit, MI, USA
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  • Kimberley S. Seok,

    1. Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA
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  • Francesca Gotsch,

    1. Perinatology Research Branch, NICHD, NIH, Bethesda, MD, and Detroit MI, USA
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  • Shali Mazaki-Tovi,

    1. Perinatology Research Branch, NICHD, NIH, Bethesda, MD, and Detroit MI, USA
    2. Department of Obstetrics and Gynecology, Wayne State University School of Medicine, Hutzel Women’s Hospital, Detroit Medical Center, Detroit, MI, USA
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  • Edi Vaisbuch,

    1. Perinatology Research Branch, NICHD, NIH, Bethesda, MD, and Detroit MI, USA
    2. Department of Obstetrics and Gynecology, Wayne State University School of Medicine, Hutzel Women’s Hospital, Detroit Medical Center, Detroit, MI, USA
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  • Katherine Sanders,

    1. Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA
    2. Department of Microbiology and Immunology, Stanford University, Stanford, CA, USA
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  • Elisabeth M. Bik,

    1. Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA
    2. Department of Microbiology and Immunology, Stanford University, Stanford, CA, USA
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  • Tinnakorn Chaiworapongsa,

    1. Perinatology Research Branch, NICHD, NIH, Bethesda, MD, and Detroit MI, USA
    2. Department of Obstetrics and Gynecology, Wayne State University School of Medicine, Hutzel Women’s Hospital, Detroit Medical Center, Detroit, MI, USA
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  • Enrique Oyarzún,

    1. Department of Obstetrics and Gynecology, P. Universidad Católica de Chile, Santiago, Chile
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  • David A. Relman

    1. Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA
    2. Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA
    3. Department of Microbiology and Immunology, Stanford University, Stanford, CA, USA
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Roberto Romero, MD, Chief, Perinatology Research Branch, 3990 John R, 4 Brush South, Detroit, MI 48201, USA.
E-mail: prbchiefstaff@med.wayne.edu
David Relman, MD, Thomas M. and Joan C. Merigan Professor, Department of Medicine – Division of Infectious Diseases, Stanford University, VA Palo Alto Health Care System 154T, Building 101, Room B4-185 3801 Miranda Avenue, Palo Alto, CA 94304, USA.
E-mail: relman@stanford.edu

Abstract

Citation DiGiulio DB, Romero R, Kusanovic JP, Gómez R, Kim CJ, Seok K, Gotsch F, Mazaki-Tovi S, Vaisbuch E, Sanders K, Bik EM, Chaiworapongsa T, Oyarzún E, Relman DA. Prevalence and diversity of microbes in the amniotic fluid, the fetal inflammatory response, and pregnancy outcome in women with preterm pre-labor rupture of membranes. Am J Reprod Immunol 2010; 64: 38–57

Problem  The role played by microbial invasion of the amniotic cavity (MIAC) in preterm pre-labor rupture of membranes (pPROM) is inadequately characterized, in part because of reliance on cultivation-based methods.

Method of study  Amniotic fluid from 204 subjects with pPROM was analyzed with both cultivation and molecular methods in a retrospective cohort study. Broad-range and group-specific polymerase chain reaction (PCR) assays targeted small subunit ribosomal DNA (rDNA), or other gene sequences, from bacteria, fungi, and archaea. Results were correlated with measurements of host inflammation, as well as pregnancy and perinatal outcomes.

Results  The prevalence of MIAC was 34% (70/204) by culture, 45% (92/204) by PCR, and 50% (101/204) by both methods combined. The number of bacterial species revealed by PCR (44 species-level phylotypes) was greater than that by culture (14 species) and included as-yet uncultivated taxa. Some taxa detected by PCR have been previously associated with the gastrointestinal tract (e.g., Coprobacillus sp.), the mouth (e.g., Rothia dentocariosa), or the vagina in the setting of bacterial vaginosis (e.g., Atopobium vaginae). The relative risk for histologic chorioamnionitis was 2.1 for a positive PCR [95% confidence interval (CI), 1.4–3.0] and 2.0 for a positive culture (95% CI, 1.4–2.7). Bacterial rDNA abundance exhibited a dose relationship with gestational age at delivery (R2 = 0.26; < 0.01). A positive PCR was associated with lower mean birthweight, and with higher rates of respiratory distress syndrome and necrotizing enterocolitis (< 0.05 for each outcome).

Conclusion  MIAC in pPROM is more common than previously recognized and is associated in some cases with uncultivated taxa, some of which are typically associated with the gastrointestinal tract. The detection of MIAC by molecular methods has clinical significance.

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