Antibody Microarray Analysis of Plasma Proteins for the Prediction of Histologic Chorioamnionitis in Women With Preterm Premature Rupture of Membranes

Abstract

We aimed to identify maternal blood biomarkers predictive of histologic chorioamnionitis (HCA) in the plasma of women with preterm premature rupture of membranes (PPROM) and to determine whether the combination of these biomarkers with conventional clinical variables can improve the prediction of HCA. This retrospective cohort study included 82 consecutive women with PPROM (23-34 gestational weeks) who delivered within 96 hours of blood sampling. A membrane-based human antibody microarray was used to analyze the plasma proteome. The validation of 5 candidate biomarkers of interest was performed by enzyme-linked immunosorbent assay (ELISA) in the final cohort (n = 82). Serum C-reactive protein (CRP) levels were measured at sampling. Seventy-nine molecules studied exhibited intergroup differences. Validation by ELISA confirmed higher levels of plasma matrix metalloproteinase-9 (MMP-9), interleukin-6 (IL-6), S100 A8/A9, and insulin-like growth factor-binding protein 1 (IGFBP-1), but not tissue inhibitor of metalloproteinase 1 (TIMP-1), in women with HCA than in women without HCA. Using a stepwise regression analysis, a combined prediction model was developed, which included the plasma MMP-9, serum CRP levels, and gestational age (area under the curve [AUC], 0.932). The AUC for this model was significantly greater than that for any single variable included in the predictive model. Protein–antibody microarray technology can be useful in identifying plasma-based predictors for HCA. This study suggests that plasma MMP-9, IL-6, IGFBP-1, and S100 A8/A9 are important noninvasive predictors for HCA in women with PPROM and that the best predictive model, which combined these biomarkers with conventional clinical factors, can significantly improve the predictability for HCA.

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

“…In the context of PPROM, IL-8 level in CVF is already consistently reported to have a significant correlation with MIAC [6,7,10,21], which is consistent with the current study. However, in the literature, CVF MMP- 9 has not yet been studied in regard to MIAC in PPROM, although it has been studied relatively extensively in AF or maternal blood [8,18,[21][22][23][24][25]. In the present study, we demonstrated that CVF MMP-9 is a useful non-invasive biomarker for MIAC in PPROM, and that the CVF level of MMP-9 was also significantly correlated with CVF IL-8 (r = 0.410).…”

“…Elevated levels of TIMP-1 have been detected in the AF in the presence of MIAC [25,36]. In contrast to the results on a sample of AF, using histologic chorioamnionitis as the outcome variable, our previous study on maternal plasma has shown that TIMP-1 levels are not significantly changed in women with PPROM [18]. At present, however, no studies have examined the changes in TIMP-1 levels in CVF associated with MIAC, and we showed that TIMP-1 levels in CVF were significantly elevated in women with PPROM and MIAC.…”

“…From each group, 500 μg of CVF samples was mixed and assayed in duplicate. The assay was conducted according to the manufacturer's instructions and as previously described [18,19]. The membranes were exposed to a radiographic film (Kodak Industrex Processor Model; Eastman Kodak Company, Rochester, NY), and the signal was detected by means of chemiluminescence image analysis (Bio-Rad Quantity 4.6.7; Bio-Rad Laboratories, Inc., Hercules, CA).…”

Maternal inflammatory markers for chorioamnionitis in preterm prelabour rupture of membranes: a systematic review and meta-analysis of diagnostic test accuracy studies

“…2, Additional file 5). In the 'Patient selection' domain, we judged 14 of the 23 studies to be at high risk of bias largely due to inappropriate exclusions such as excluding women based on duration after PPROM [35,38], not explicitly excluding women with clinical features of chorioamnionitis at the time of PPROM or at the time of admission [26,27,31,34,38,40], basing exclusions on availability or ability to perform other tests [31,40,45], excluding women due to missing data [34,35,50] and excluding women with common conditions and complications of pregnancy that often coexist with PPROM [32,36,39,40]. In the 'Index test' domain, all tests were considered to be 'blinded' because maternal blood was collected before delivery and assessed on automated assays.…”

“…Six studies specified that clinical features of chorioamnionitis were an indication for delivery [26,28,29,36,44,49]. According to the definitions of reference standard provided, 11 studies [26,28,31,35,36,[40][41][42][43][44]50] reported the index test against a reference standard of HCA and/ or funisitis, 10 [27, 29, 30, 32, 34, 39, 45, 46, 48, 49] reported HCA alone and 2 studies [33,38] reported funisitis alone. Characteristics of included studies are outlined in Table 1.…”