Skip to main content

Advertisement

SpringerLink
Log in

Cervicovaginal Fluid Protein Microarray for Detection of Microbial Invasion of the Amniotic Cavity in Preterm Labor

  • Original Article
  • Published:
Reproductive Sciences Aims and scope Submit manuscript

Abstract

We aimed to identify cervicovaginal fluid (CVF) biomarkers that can detect microbial invasion of the amniotic cavity (MIAC) in women with preterm labor (PTL) with an antibody microarray and to develop the best combined model for detection of MIAC using these biomarkers in combination with conventional clinical variables. This retrospective cohort study included 168 singleton pregnant women with PTL (23–34 weeks) who underwent amniocentesis. AF was cultured, and CVF samples were obtained at the time of amniocentesis. An antibody microarray was used to analyze the CVF proteome (n = 40). The validation of four candidate biomarkers of interest was performed by enzyme-linked immunosorbent assay (ELISA) in the final cohort (n = 168). For comparison with candidate markers, CVF IL-6 concentration was also measured. Twenty-seven molecules studied exhibited intergroup differences. Validation by ELISA confirmed significantly higher levels of CVF DKK3, M-CSF, and TIMP-1, but not of IGFBP-2, independent of gestational age, in CVF of women with MIAC. The area under the curve (AUC) of DKK3, M-CSF, and TIMP-1 from CVF was not significantly different from the AUC of IL-6 from CVF for detecting MIAC in women with PTL. By using a stepwise regression analysis, a combined detection model was developed, which included the CVF M-CSF, TIMP-1, and gestational age at sampling (AUC = 0.823). An antibody microarray identified useful biomarkers (DKK3, M-CSF, and TIMP-1) in CVF for detection of MIAC, and a combined model including these biomarkers and gestational age can accurately detect MIAC in women with PTL.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Slattery MM, Morrison JJ. Preterm delivery. Lancet. 2002;360(9344):1489–97.

    Article  Google Scholar 

  2. Zeitlin J, Draper ES, Kollee L, et al. Differences in rates and short-term outcome of live births before 32 weeks of gestation in Europe in 2003: results from the MOSAIC cohort. Pediatrics. 2008;121(4):e936–44.

    Article  Google Scholar 

  3. Goncalves LF, Chaiworapongsa T, Romero R. Intrauterine infection and prematurity. Ment Retard Dev Disabil Res Rev. 2002;8(1):3–13.

    Article  Google Scholar 

  4. Bashiri A, Burstein E, Mazor M. Cerebral palsy and fetal inflammatory response syndrome: a review. J Perinat Med. 2006;34(1):5–12.

    Article  Google Scholar 

  5. Yoon BH, Chang JW, Romero R. Isolation of Ureaplasma urealyticum from the amniotic cavity and adverse outcome in preterm labor. Obstet Gynecol. 1998;92(1):77–82.

    Article  CAS  Google Scholar 

  6. Combs CA, Gravett M, Garite TJ, et al. Amniotic fluid infection, inflammation, and colonization in preterm labor with intact membranes. Am J Obstet Gynecol. 2014;210(2):125 e121–15.

    Article  Google Scholar 

  7. Jung EY, Choi BY, Rhee J, Park J, Cho SH, Park KH. Relation between amniotic fluid infection or cytokine levels and hearing screen failure in infants at 32 wk gestation or less. Pediatr Res. 2017;81(2):349–55.

    Article  CAS  Google Scholar 

  8. Di Quinzio MK, Georgiou HM, Holdsworth-Carson SJ, et al. Proteomic analysis of human cervico-vaginal fluid displays differential protein expression in association with labor onset at term. J Proteome Res. 2008;7(5):1916–21.

    Article  Google Scholar 

  9. Holst RM, Hagberg H, Wennerholm UB, Skogstrand K, Thorsen P, Jacobsson B. Prediction of microbial invasion of the amniotic cavity in women with preterm labour: analysis of multiple proteins in amniotic and cervical fluids. BJOG. 2011;118(2):240–9.

    Article  Google Scholar 

  10. Rizzo G, Capponi A, Rinaldo D, Tedeschi D, Arduini D, Romanini C. Interleukin-6 concentrations in cervical secretions identify microbial invasion of the amniotic cavity in patients with preterm labor and intact membranes. Am J Obstet Gynecol. 1996;175(4 Pt 1):812–7.

    Article  CAS  Google Scholar 

  11. Combs CA, Garite TJ, Lapidus JA, et al. Detection of microbial invasion of the amniotic cavity by analysis of cervicovaginal proteins in women with preterm labor and intact membranes. Am J Obstet Gynecol. 2015;212(4):482 e481–12.

    Article  Google Scholar 

  12. Romero R, Yoon BH, Mazor M, Gomez R, Diamond MP, Kenney JS, et al. The diagnostic and prognostic value of amniotic fluid white blood cell count, glucose, interleukin-6, and gram stain in patients with preterm labor and intact membranes. Am J Obstet Gynecol. 1993;169(4):805–16.

    Article  CAS  Google Scholar 

  13. Sutandy FX, Qian J, Chen CS, Zhu H. Overview of protein microarrays. Curr Protoc Protein Sci. 2013;Chapter 27:Unit 27 21.

  14. Haab BB. Antibody arrays in cancer research. Mol Cell Proteomics. 2005;4(4):377–83.

    Article  CAS  Google Scholar 

  15. Lee SY, Park KH, Jeong EH, Oh KJ, Ryu A, Kim A. Intra-amniotic infection/inflammation as a risk factor for subsequent ruptured membranes after clinically indicated amniocentesis in preterm labor. J Korean Med Sci. 2013;28(8):1226–32.

    Article  CAS  Google Scholar 

  16. Jung EY, Park KH, Han BR, Cho SH, Ryu A. Measurement of interleukin 8 in Cervicovaginal fluid in women with preterm premature rupture of membranes: a comparison of amniotic fluid samples. Reprod Sci. 2017;24(1):142–7.

    Article  CAS  Google Scholar 

  17. Cha DM, Woo SJ, Kim HJ, Lee C, Park KH. Comparative analysis of aqueous humor cytokine levels between patients with exudative age-related macular degeneration and normal controls. Invest Ophthalmol Vis Sci. 2013;54(10):7038–44.

    Article  CAS  Google Scholar 

  18. Park JW, Park KH, Lee JE, Kim YM, Lee SJ, Cheon DH. Antibody microarray analysis of plasma proteins for the prediction of histologic Chorioamnionitis in women with preterm premature rupture of membranes. Reprod Sci. 2019;1933719119828043.

  19. DeLong ER, DeLong DM, Clarke-Pearson DL. Comparing the areas under two or more correlated receiver operating characteristic curves: a nonparametric approach. Biometrics. 1988;44(3):837–45.

    Article  CAS  Google Scholar 

  20. Jung EY, Park JW, Ryu A, Lee SY, Cho SH, Park KH. Prediction of impending preterm delivery based on sonographic cervical length and different cytokine levels in cervicovaginal fluid in preterm labor. J Obstet Gynaecol Res. 2016;42(2):158–65.

    Article  CAS  Google Scholar 

  21. Liong S, Di Quinzio MK, Fleming G, Permezel M, Rice GE, Georgiou HM. New biomarkers for the prediction of spontaneous preterm labour in symptomatic pregnant women: a comparison with fetal fibronectin. BJOG. 2015;122(3):370–9.

    Article  CAS  Google Scholar 

  22. Marvin KW, Keelan JA, Coleman MA, McCowan LM, Zhou RL, Mitchell MD. Intercellular adhesion molecule-1 (ICAM-1) in cervicovaginal fluid of women presenting with preterm labor: predictive value for preterm delivery. Am J Reprod Immunol. 2000;43(5):264–71.

    Article  CAS  Google Scholar 

  23. Parker J, Bell R, Brennecke S. Fetal fibronectin in the cervicovaginal fluid of women with threatened preterm labour as a predictor of delivery before 34 weeks' gestation. Aust N Z J Obstet Gynaecol. 1995;35(3):257–61.

    Article  CAS  Google Scholar 

  24. Jacobsson B, Holst RM, Mattsby-Baltzer I, Nikolaitchouk N, Wennerholm UB, Hagberg H. Interleukin-18 in cervical mucus and amniotic fluid: relationship to microbial invasion of the amniotic fluid, intra-amniotic inflammation and preterm delivery. BJOG. 2003;110(6):598–603.

    Article  CAS  Google Scholar 

  25. Park JW, Park KH, Lee SY. Noninvasive prediction of intra-amniotic infection and/or inflammation in women with preterm labor: various cytokines in cervicovaginal fluid. Reprod Sci. 2013;20(3):262–8.

    Article  Google Scholar 

  26. Bharti K, Gasper M, Ou J, Brucato M, Clore-Gronenborn K, Pickel J, et al. A regulatory loop involving PAX6, MITF, and WNT signaling controls retinal pigment epithelium development. PLoS Genet. 2012;8(7):e1002757.

    Article  CAS  Google Scholar 

  27. Yoo HN, Park KH, Jung EY, Kim YM, Kook SY, Jeon SJ. Non-invasive prediction of preterm birth in women with cervical insufficiency or an asymptomatic short cervix (</=25 mm) by measurement of biomarkers in the cervicovaginal fluid. PLoS One. 2017;12(7):e0180878.

    Article  Google Scholar 

  28. Lee SE, Romero R, Park CW, Jun JK, Yoon BH. The frequency and significance of intraamniotic inflammation in patients with cervical insufficiency. Am J Obstet Gynecol. 2008;198(6):633.e1–8.

    Article  Google Scholar 

  29. Chitu V, Stanley ER. Colony-stimulating factor-1 in immunity and inflammation. Curr Opin Immunol. 2006;18(1):39–48.

    Article  CAS  Google Scholar 

  30. Piccinni MP, Scaletti C, Vultaggio A, Maggi E, Romagnani S. Defective production of LIF, M-CSF and Th2-type cytokines by T cells at fetomaternal interface is associated with pregnancy loss. J Reprod Immunol. 2001;52(1–2):35–43.

    Article  CAS  Google Scholar 

  31. Eckmann-Scholz C, Wilke C, Acil Y, Alkatout I, Salmassi A. Macrophage colony-stimulating factor (M-CSF) in first trimester maternal serum: correlation with pathologic pregnancy outcome. Arch Gynecol Obstet. 2016;293(6):1213–7.

    Article  CAS  Google Scholar 

  32. Hayashi M, Zhu K, Sagesaka T, Fukasawa I, Inaba N. Elevation of amniotic fluid macrophage colony-stimulating factor in normotensive pregnancies that delivered small-for-gestational-age infants. Am J Reprod Immunol. 2007;57(6):488–94.

    Article  CAS  Google Scholar 

  33. Hayashi M, Hombo Y, Shibazaki M, Nakajima A, Inaba N. Elevation of macrophage colony-stimulating factor in amniotic fluid at late stage of normal pregnancy. Am J Reprod Immunol. 2006;55(3):226–31.

    Article  CAS  Google Scholar 

  34. Sundrani D, Narang A, Mehendale S, Joshi S, Chavan-Gautam P. Investigating the expression of MMPs and TIMPs in preterm placenta and role of CpG methylation in regulating MMP-9 expression. IUBMB Life. 2017;69(12):985–93.

    Article  CAS  Google Scholar 

  35. Clark IM, Morrison JJ, Hackett GA, Powell EK, Cawston TE, Smith SK. Tissue inhibitor of metalloproteinases: serum levels during pregnancy and labor, term and preterm. Obstet Gynecol. 1994;83(4):532–7.

    Article  CAS  Google Scholar 

  36. Geng J, Huang C, Jiang S. Roles and regulation of the matrix metalloproteinase system in parturition. Mol Reprod Dev. 2016;83(4):276–86.

    Article  CAS  Google Scholar 

  37. Park JW, Park KH, Kook SY, Jung YM, Kim YM. Immune biomarkers in maternal plasma to identify histologic chorioamnionitis in women with preterm labor. Arch Gynecol Obstet. 2019;299(3):725–32.

    Article  CAS  Google Scholar 

  38. Oh KJ, Lee SE, Jung H, Kim G, Romero R, Yoon BH. Detection of ureaplasmas by the polymerase chain reaction in the amniotic fluid of patients with cervical insufficiency. J Perinat Med. 2010, 38;(3):261–8.

Download references

Funding

This study was supported by the Seoul National University Bundang Hospital Research Fund (Grant No. 13–2017-004) and 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 & Welfare, Republic of Korea (grant number: HI18C0063).

Author information

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, South Korea

    Seung Mi Lee, Kyo Hoon Park, So Yeon Kim & Subeen Hong

  2. Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, 82-ro 173 Beon-gil, Seongnamsi, Kyeonggido, Gumi, 463-707, South Korea

    Kyo Hoon Park, So Yeon Kim, Yu Mi Kim & Subeen Hong

  3. Department of Laboratory Medicine, Seoul National University Boramae Hospital, 20 Boramae-ro 5-gil, Sindaebang-dong, Dongjak-gu, Seoul, South Korea

    Sue Shin

Authors
  1. Seung Mi Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kyo Hoon Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. So Yeon Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yu Mi Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Subeen Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sue Shin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kyo Hoon Park.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic Supplementary Material

ESM 1

(DOCX 31 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lee, S.M., Park, K.H., Kim, S.Y. et al. Cervicovaginal Fluid Protein Microarray for Detection of Microbial Invasion of the Amniotic Cavity in Preterm Labor. Reprod. Sci. 27, 713–721 (2020). https://doi.org/10.1007/s43032-019-00077-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s43032-019-00077-6

Keywords

Search

Navigation