Background: Escherichia coli early-onset sepsis (EOS) is an important cause of mortality and morbidity in neonates, corresponding to the increasing prevalence in the community, the carriage of E. Coli in pregnant women has also been increased and vaginal E. Coli carriage predisposes newborns to colonization during the birth process. Aim: Vertical transmission of E.coli colonization in healthy newborn, born to mothers with and without colonization Methods: This case control study was conducted in the Department of Paediatrics, Services Hospital and Jinnah hospital, Lahore from January to June 2015 (ongoing). The mothers were subjected to vaginal swab and anterior nares swab for E. coli within 6 hours prior to plan delivery. The nasal swab for culture from anterior nares of newborn was obtained within 1 hour of birth. Samples were cultured in Paediatric Microbiology laboratory. Data were analyzed and compared. Results: Out of total, 80 mothers and their newborns, 10 had E. coli colonization. Conclusion: E.coli colonization was detected in 12.5% of infants.