Aim: This study was carried out to determine the vaginal bacteriological pattern along with their antibiotic susceptibility pattern in pregnant women with preterm prelabor rupture of membranes (PPROM) and to compare them with those without PPROM. The association of PPROM with neonatal and maternal complications was also studied. Materials and methods: This is an observational comparative study conducted in the year 2020 at a tertiary care teaching hospital located in Puducherry, India. The study included singleton pregnancies between 28 0/7 and 36 6/7 weeks of gestation with PPROM along with gestational age matched women without PPROM. A high vaginal swab was taken from all the candidates and sent for aerobic and anaerobic cultures. Organisms isolated along with their antibiotic susceptibility were noted and compared between the two groups. The software used was Statistical Package for the Social Sciences (SPSS) software, version 17.0 (IBM, Armonk, NY, USA). Results: This study demonstrated an overall incidence of PPROM of 2.8%. A positive culture rate of 35.2 and 9.3% was found among pregnant women with and without PPROM, which was found to be statistically significant. The commonest organism isolated was Escherichia coli (52.6%) which was 100% sensitive to imipenem and meropenem and 90% to cefoperazone–sulbactam and was found to be resistant to ampicillin in 90% of cases. Neonatal complications such as respiratory distress and sepsis were found to be significantly higher in the PPROM group than in the comparison group. Conclusion: There is no change in the vaginal microbiota in the last two decades, but the sensitivity pattern of antibiotics has changed considerably from the commonly used antibiotics to higher-generation antibiotics. This indicates indiscriminate use of antibiotics which has resulted in antibiotic resistance. This could be the probable reason for the increase in neonatal and maternal morbidity in PPROM. Clinical significance: The vaginal microbiological flora and the antibiotic susceptibility pattern of the population in this area was studied and a significant change was noted over the past years which will help in formulating newer and better treatment protocols in the future which would help in reducing neonatal and maternal morbidities.