Protein microarray analysis of amniotic fluid proteins associated with spontaneous preterm birth in women with preterm labor

Problem

To identify proteins present in the amniotic fluid (AF) that could be associated with spontaneous preterm birth (SPTB; delivery < 7 days) in women with preterm labor (PTL).

Method of study

First, the AF proteome of 20 women with PTL and SPTB was compared with that of 20 matched women with term deliveries using an antibody microarray. Next, nine identified candidate biomarkers of SPTB were further validated in 267 singleton pregnant women with PTL who underwent amniocentesis at 26–33 weeks of gestation using ELISA, and whether the degree of expression of these proteins was associated with the risk severity for subsequent SPTB was retrospectively assessed.

Results

Of the 507 proteins evaluated in the microarray analysis, 27 displayed significant intergroup differences. In particular, ELISA quantification confirmed that the expression of EN-RAGE, IL-6, IL-8, IP-10, lipocalin-2, MMP-8, MMP-9, S100 A8/A9, and TNFR2 were all increased in the AF of women spontaneously delivering within 7 days of sampling compared with those delivering after 7 days. Moreover, the odds of SPTB within 7 days, even upon adjusting for confounders, tended to significantly increase with each increasing quartile of baseline AF levels of each protein (P-value for trend < 0.05).

Conclusion

Nine AF proteins were found to be independently associated with higher risk of subsequent SPTB in women with PTL, all of which were immune-, inflammation-, and extracellular matrix-related proteins. Moreover, risk severity for this subsequent SPTB is closely related to the degree of expression of each of these proteins.

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