BACKGROUND: Premature rupture of membranes (PROM) is a common pregnancy complication and is associated with significant risks of fetal and maternal morbidity and mortality. However, accurate diagnosis of PROM remains a frequent clinical problem in obstetrics especially in the developing countries. Accurate and timely diagnosis is therefore important because failure of it can lead to failure to implement appropriate obstetrics measures. OBJECTIVES: This study explored the hypothesis that AmniSure test will provide more accurate results on the state of fetal membranes than the combined use of the standard diagnostic techniques of pooling of fluids, ferning, and nitrazine testing in sensitivity and specificity, with the potential to serve as a gold standard for diagnosing PROM. This study determined the accuracy of monoclonal antibody (AmniSure) test in comparison with nitrazine test, fern test and pooling of fluid in the diagnosis of PROM among pregnant women in Nnamdi Azikiwe University Teaching Hospital, Nnewi, South-east Nigeria. METHODOLOGY: A prospective observational study was performed in consecutive women with signs or symptoms of rupture of membranes. Initial evaluation included both the standard clinical assessment for rupture of membranes and AmniSure test. Rupture of membranes was diagnosed if fluid was seen leaking from the cervical os or if two of the following three conditions were present: pooling of fluid, positive nitrazine test, or ferning. Rupture of membranes was diagnosed definitively on review of the medical records after delivery. Descriptive analysis of the result was done using the Epi- info 2008 version 3.5.1. Analyses were performed using chi-square (χ2) tests where appropriate. A probability value of <0.05 was considered significant.