Background: Premature or pre-labour rupture of membranes is a known important contributor to maternal and perinatal morbidity and mortality. The increasing concentration of creatinine and urea in the amniotic fluid throughout normal pregnancy make these biomarkers invaluable in the diagnosis of PROM in equivocal cases. Aim: The aim of this study was to evaluate the value of vaginal fluid levels of creatinine and urea in the diagnosis of premature rupture of membranes (PROM). Materials and methods: A total of 195 pregnant women in three equal groups who met the inclusion criteria participated in this prospective study conducted in the department of Obstetrics and Gynaecology, Irrua Specialist Teaching Hospital Irrua Edo State. Group I were women who had rupture of membrane confirmed by visual pooling and nitrazine positive test. Group II were patients who complained of vaginal fluid leakage in whom PROM was suspected but unconfirmed by vaginal pooling and had negative nitrazine test. Group III consisted of normal pregnant women without complaint. All participants were sampled for cervicovaginal fluid urea and creatinine by speculum examination. One-way ANOVA, chi-square, correlation coefficient and receiver operator characteristics (ROC) curve analysis were used.