Objective: To determine whether various immune-related proteins in cervicovaginal fluid (CVF), alone or in combination with clinical risk factors, can predict intra-amniotic infection and/or inflammation (IAI) in women with preterm premature rupture of membranes (PPROM) and to compare predictive ability of these biomarkers with that of amniotic fluid (AF) white blood cell (WBC) using invasive amniocentesis. Methods: This retrospective cohort study included 99 consecutive singleton pregnant women (23.0-33.6 gestational weeks) with PPROM who underwent amniocentesis. The amniotic fluid (AF) was cultured, and the white blood cell (WBC) count was determined. The stored CVF samples were assayed for complement C3a and C5a, TIMP-1, MMP-9, and M-CSF by using ELISA kits. Serum C-reactive protein (CRP) level was measured at the time of sampling. The primary outcome measures were IAI [defined as a positive AF culture and/or an elevated AF IL-6 level (2.6 ng/mL)]. Results: The overall prevalence of IAI was 50.5% (50/99). The CVF levels of C3a, C5a, TIMP-1, MMP-9, and M-CSF were significantly higher in women with IAI. The women with IAI had a significantly lower gestational age at sampling and a higher level of serum CRP. Using a stepwise regression analysis, a combined prediction model was developed, which included the CVF MMP-9 and serum CRP [area under the curve (AUC), 0.827]. The AUC for this model was significantly higher than either parameter retained in this model, but no differences were observed between the AUC of this model based on non-invasive variables, and AF WBC. Conclusion: This study suggests that TIMP-1, MMP-9, M-CSF, C3a and C5a in the CVF may be useful as new non-invasive predictors of IAI in women with PPROM and that the best predictive model, which combined these biomarkers with conventional clinical factors, can significantly improve the predictability for IAI.