Widespread microbial invasion of the chorioamniotic membranes is a consequence and not a cause of intra-amniotic infection

Mi Jeong Kim, Roberto Romero, Maria Teresa Gervasi, Jung Sun Kim, Wonsuk Yoo, Deug Chan Lee, Pooja Mittal, Offer Erez, Juan Pedro Kusanovic, Sonia S. Hassan, Chong Jai Kim

Research output: Contribution to journalArticle

85 Citations (Scopus)

Abstract

Acute chorioamnionitis is a response to amniotic fluid (AF) infection. However, it remains unclear whether substantial bacterial propagation in the chorioamniotic membranes (CAMs) precedes microbial invasion of the amniotic cavity (MIAC), which is inconsistent with characteristic amniotropic neutrophil migration in acute chorioamnionitis. This study was performed to determine whether CAMs have widespread bacterial infection during MIAC and whether bacteria normally colonize CAMs. AF pellets and CAMs from the following groups were studied: group 1, patients with positive (n18) or negative (n22) AF cultures; group 2, patients with or without acute chorioamnionitis in which the amnion and chorion were studied separately (n60); and group 3, patients at term who underwent a cesarean delivery (n30). SYTO 9/propidium iodide fluorescent staining and fluorescent in situ hybridization for 16S rRNA were performed. Real-time quantitative PCR for 16S rDNA and PCR for genital mycoplasmas were also conducted. Bacteria were more frequently detected in AF than in CAMs of patients with positive AF culture (100 vs. 33%; P0.0001). Bacteria were detected more frequently in CAMs as the severity of chorioamnionitis increased (P0.01). The median 16S rRNA gene copy number in the amnion was significantly greater than in the chorion (group 2; P0.0001). Bacteria were not detected in CAMs or AF in women at term before labor (group 3). A fraction of patients with chorioamnionitis or MIAC did not have bacteria in CAMs. Collectively, the findings herein indicate that MIAC does not follow widespread infection of CAMs, but precedes it. We propose a model of MIAC: the initial stage is intra-amniotic bacterial invasion through a discrete region of the CAMs, followed by intra-amniotic proliferation, and bacterial invasion of CAMs primarily extends from the amniotic fluid. This study emphasizes the importance of assessing the intra-amniotic compartment for diagnosis and treatment of preterm birth.

Original languageEnglish (US)
Pages (from-to)924-936
Number of pages13
JournalLaboratory Investigation
Volume89
Issue number8
DOIs
StatePublished - Aug 1 2009

Fingerprint

Amniotic Fluid
Chorioamnionitis
Membranes
Infection
Bacteria
Chorion
Amnion
Gene Dosage
Propidium
Mycoplasma
Premature Birth
Ribosomal DNA
Fluorescence In Situ Hybridization
rRNA Genes
Bacterial Infections
Real-Time Polymerase Chain Reaction
Neutrophils
Staining and Labeling
Polymerase Chain Reaction

Keywords

  • 16S rRNA
  • Amniotic fluid
  • Chorioamnionitis
  • Fluorescent in situ hybridization
  • Intra-amniotic infection
  • Polymerase chain reaction

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Biology
  • Cell Biology

Cite this

Widespread microbial invasion of the chorioamniotic membranes is a consequence and not a cause of intra-amniotic infection. / Kim, Mi Jeong; Romero, Roberto; Gervasi, Maria Teresa; Kim, Jung Sun; Yoo, Wonsuk; Lee, Deug Chan; Mittal, Pooja; Erez, Offer; Kusanovic, Juan Pedro; Hassan, Sonia S.; Kim, Chong Jai.

In: Laboratory Investigation, Vol. 89, No. 8, 01.08.2009, p. 924-936.

Research output: Contribution to journalArticle

Kim, MJ, Romero, R, Gervasi, MT, Kim, JS, Yoo, W, Lee, DC, Mittal, P, Erez, O, Kusanovic, JP, Hassan, SS & Kim, CJ 2009, 'Widespread microbial invasion of the chorioamniotic membranes is a consequence and not a cause of intra-amniotic infection', Laboratory Investigation, vol. 89, no. 8, pp. 924-936. https://doi.org/10.1038/labinvest.2009.49
Kim, Mi Jeong ; Romero, Roberto ; Gervasi, Maria Teresa ; Kim, Jung Sun ; Yoo, Wonsuk ; Lee, Deug Chan ; Mittal, Pooja ; Erez, Offer ; Kusanovic, Juan Pedro ; Hassan, Sonia S. ; Kim, Chong Jai. / Widespread microbial invasion of the chorioamniotic membranes is a consequence and not a cause of intra-amniotic infection. In: Laboratory Investigation. 2009 ; Vol. 89, No. 8. pp. 924-936.
@article{8f3752bb331c4f6fa54ed1c23f41da20,
title = "Widespread microbial invasion of the chorioamniotic membranes is a consequence and not a cause of intra-amniotic infection",
abstract = "Acute chorioamnionitis is a response to amniotic fluid (AF) infection. However, it remains unclear whether substantial bacterial propagation in the chorioamniotic membranes (CAMs) precedes microbial invasion of the amniotic cavity (MIAC), which is inconsistent with characteristic amniotropic neutrophil migration in acute chorioamnionitis. This study was performed to determine whether CAMs have widespread bacterial infection during MIAC and whether bacteria normally colonize CAMs. AF pellets and CAMs from the following groups were studied: group 1, patients with positive (n18) or negative (n22) AF cultures; group 2, patients with or without acute chorioamnionitis in which the amnion and chorion were studied separately (n60); and group 3, patients at term who underwent a cesarean delivery (n30). SYTO 9/propidium iodide fluorescent staining and fluorescent in situ hybridization for 16S rRNA were performed. Real-time quantitative PCR for 16S rDNA and PCR for genital mycoplasmas were also conducted. Bacteria were more frequently detected in AF than in CAMs of patients with positive AF culture (100 vs. 33{\%}; P0.0001). Bacteria were detected more frequently in CAMs as the severity of chorioamnionitis increased (P0.01). The median 16S rRNA gene copy number in the amnion was significantly greater than in the chorion (group 2; P0.0001). Bacteria were not detected in CAMs or AF in women at term before labor (group 3). A fraction of patients with chorioamnionitis or MIAC did not have bacteria in CAMs. Collectively, the findings herein indicate that MIAC does not follow widespread infection of CAMs, but precedes it. We propose a model of MIAC: the initial stage is intra-amniotic bacterial invasion through a discrete region of the CAMs, followed by intra-amniotic proliferation, and bacterial invasion of CAMs primarily extends from the amniotic fluid. This study emphasizes the importance of assessing the intra-amniotic compartment for diagnosis and treatment of preterm birth.",
keywords = "16S rRNA, Amniotic fluid, Chorioamnionitis, Fluorescent in situ hybridization, Intra-amniotic infection, Polymerase chain reaction",
author = "Kim, {Mi Jeong} and Roberto Romero and Gervasi, {Maria Teresa} and Kim, {Jung Sun} and Wonsuk Yoo and Lee, {Deug Chan} and Pooja Mittal and Offer Erez and Kusanovic, {Juan Pedro} and Hassan, {Sonia S.} and Kim, {Chong Jai}",
year = "2009",
month = "8",
day = "1",
doi = "10.1038/labinvest.2009.49",
language = "English (US)",
volume = "89",
pages = "924--936",
journal = "Laboratory Investigation",
issn = "0023-6837",
publisher = "Nature Publishing Group",
number = "8",

}

TY - JOUR

T1 - Widespread microbial invasion of the chorioamniotic membranes is a consequence and not a cause of intra-amniotic infection

AU - Kim, Mi Jeong

AU - Romero, Roberto

AU - Gervasi, Maria Teresa

AU - Kim, Jung Sun

AU - Yoo, Wonsuk

AU - Lee, Deug Chan

AU - Mittal, Pooja

AU - Erez, Offer

AU - Kusanovic, Juan Pedro

AU - Hassan, Sonia S.

AU - Kim, Chong Jai

PY - 2009/8/1

Y1 - 2009/8/1

N2 - Acute chorioamnionitis is a response to amniotic fluid (AF) infection. However, it remains unclear whether substantial bacterial propagation in the chorioamniotic membranes (CAMs) precedes microbial invasion of the amniotic cavity (MIAC), which is inconsistent with characteristic amniotropic neutrophil migration in acute chorioamnionitis. This study was performed to determine whether CAMs have widespread bacterial infection during MIAC and whether bacteria normally colonize CAMs. AF pellets and CAMs from the following groups were studied: group 1, patients with positive (n18) or negative (n22) AF cultures; group 2, patients with or without acute chorioamnionitis in which the amnion and chorion were studied separately (n60); and group 3, patients at term who underwent a cesarean delivery (n30). SYTO 9/propidium iodide fluorescent staining and fluorescent in situ hybridization for 16S rRNA were performed. Real-time quantitative PCR for 16S rDNA and PCR for genital mycoplasmas were also conducted. Bacteria were more frequently detected in AF than in CAMs of patients with positive AF culture (100 vs. 33%; P0.0001). Bacteria were detected more frequently in CAMs as the severity of chorioamnionitis increased (P0.01). The median 16S rRNA gene copy number in the amnion was significantly greater than in the chorion (group 2; P0.0001). Bacteria were not detected in CAMs or AF in women at term before labor (group 3). A fraction of patients with chorioamnionitis or MIAC did not have bacteria in CAMs. Collectively, the findings herein indicate that MIAC does not follow widespread infection of CAMs, but precedes it. We propose a model of MIAC: the initial stage is intra-amniotic bacterial invasion through a discrete region of the CAMs, followed by intra-amniotic proliferation, and bacterial invasion of CAMs primarily extends from the amniotic fluid. This study emphasizes the importance of assessing the intra-amniotic compartment for diagnosis and treatment of preterm birth.

AB - Acute chorioamnionitis is a response to amniotic fluid (AF) infection. However, it remains unclear whether substantial bacterial propagation in the chorioamniotic membranes (CAMs) precedes microbial invasion of the amniotic cavity (MIAC), which is inconsistent with characteristic amniotropic neutrophil migration in acute chorioamnionitis. This study was performed to determine whether CAMs have widespread bacterial infection during MIAC and whether bacteria normally colonize CAMs. AF pellets and CAMs from the following groups were studied: group 1, patients with positive (n18) or negative (n22) AF cultures; group 2, patients with or without acute chorioamnionitis in which the amnion and chorion were studied separately (n60); and group 3, patients at term who underwent a cesarean delivery (n30). SYTO 9/propidium iodide fluorescent staining and fluorescent in situ hybridization for 16S rRNA were performed. Real-time quantitative PCR for 16S rDNA and PCR for genital mycoplasmas were also conducted. Bacteria were more frequently detected in AF than in CAMs of patients with positive AF culture (100 vs. 33%; P0.0001). Bacteria were detected more frequently in CAMs as the severity of chorioamnionitis increased (P0.01). The median 16S rRNA gene copy number in the amnion was significantly greater than in the chorion (group 2; P0.0001). Bacteria were not detected in CAMs or AF in women at term before labor (group 3). A fraction of patients with chorioamnionitis or MIAC did not have bacteria in CAMs. Collectively, the findings herein indicate that MIAC does not follow widespread infection of CAMs, but precedes it. We propose a model of MIAC: the initial stage is intra-amniotic bacterial invasion through a discrete region of the CAMs, followed by intra-amniotic proliferation, and bacterial invasion of CAMs primarily extends from the amniotic fluid. This study emphasizes the importance of assessing the intra-amniotic compartment for diagnosis and treatment of preterm birth.

KW - 16S rRNA

KW - Amniotic fluid

KW - Chorioamnionitis

KW - Fluorescent in situ hybridization

KW - Intra-amniotic infection

KW - Polymerase chain reaction

UR - http://www.scopus.com/inward/record.url?scp=68149164687&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=68149164687&partnerID=8YFLogxK

U2 - 10.1038/labinvest.2009.49

DO - 10.1038/labinvest.2009.49

M3 - Article

VL - 89

SP - 924

EP - 936

JO - Laboratory Investigation

JF - Laboratory Investigation

SN - 0023-6837

IS - 8

ER -