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Identification of Cultivable Bacteria in Amniotic Fluid Using Cervicovaginal Fluid Protein Microarray in Preterm Premature Rupture of Membranes

  • Seung Mi Lee
  • Kyo Hoon ParkEmail author
  • Subeen Hong
  • Yu Mi Kim
  • Ye Hyon Park
  • Young Eun Lee
  • Se Jeong Jeon
Original Article
  • 3 Downloads

Abstract

We aimed to identify cervicovaginal fluid (CVF) protein biomarkers of microbial invasion of the amniotic cavity (MIAC) in women with preterm premature rupture of membranes (PPROM), using an antibody microarray. This retrospective cohort study included 99 consecutive women with singleton pregnancies and PPROM (23–33 weeks) who underwent amniocentesis and who gave CVF samples. CVF proteomes from the MIAC (n = 20) versus non-MIAC groups (n = 20) were comparatively profiled by an antibody microarray using a nested case-control study design. The seven candidate biomarkers of interest were validated in the total cohort (n = 99) by enzyme-linked immunosorbent assays (ELISA). For comparison with candidate markers, amniotic fluid (AF) white blood cell (WBC) count was also measured. The primary outcome measure was MIAC (defined as positive AF culture). Thirty of the proteins studied exhibited significant intergroup differences. Measurements of the total cohort with ELISA confirmed a significant increase in the levels of CVF IL-8, lipocalin-2, MIP-1α, MMP-9, and TIMP-1 in women with MIAC, independent of gestational age at sampling. A combined, non-invasive model was developed by using a stepwise regression procedure, which included CVF IL-8 and CVF MMP-9 (area under the curve [AUC] = 0.763), and this AUC was comparable with the AUC of AF WBC. Using protein–antibody microarray technology, we found several novel, independent, non-invasive biomarkers to identify MIAC in women with PPROM: IL-8, lipocalin-2, MIP-1α, MMP-9, and TIMP-1. Furthermore, the combined non-invasive model (IL-8 and MMP-9) was a useful independent predictor for MIAC with good discriminatory power, similar to AF WBC count.

Keywords

Antibody microarray Biomarker Cervicovaginal fluid Microbial invasion of the amniotic cavity Preterm premature rupture of membranes 

Notes

Funding Information

This study was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health and Welfare, Republic of Korea (grant number: HI18C0063).

Compliance with Ethical Standards

At enrollment, patients with PPROM provided consent for the collection and use of CVF samples and for the use of their clinical information for research purposes. This retrospective cohort study was approved by the Ethics Committee at Seoul National University Bundang Hospital, Seongnamsi, Republic of Korea (IRB no. B-1105/128-102).

Conflict of Interest

The authors declare that there is no conflict of interest.

Supplementary material

43032_2020_143_MOESM1_ESM.docx (20 kb)
ESM 1 (DOCX 20.1 kb)
43032_2020_143_MOESM2_ESM.docx (24 kb)
Table S1 (DOCX 24 kb)

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Copyright information

© Society for Reproductive Investigation 2020

Authors and Affiliations

  • Seung Mi Lee
    • 1
    • 2
  • Kyo Hoon Park
    • 1
    • 3
    Email author
  • Subeen Hong
    • 1
    • 3
  • Yu Mi Kim
    • 3
  • Ye Hyon Park
    • 3
  • Young Eun Lee
    • 3
  • Se Jeong Jeon
    • 1
  1. 1.Department of Obstetrics and GynecologySeoul National University College of MedicineSeoulSouth Korea
  2. 2.Department of Obstetrics and GynecologySeoul National University HospitalSeoulSouth Korea
  3. 3.Department of Obstetrics and GynecologySeoul National University Bundang HospitalSeongnamsiSouth Korea

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