This prevalence study was conducted to compare the counts, types and antimicrobial resistance profiles of bacterial flora on the hands of individuals in the community to that of nurses at a nearby university teaching hospital, with an intense hand hygiene regimen. Hand cultures were obtained from 204 individuals during a home visit and 119 nurses in two neonatal intensive care units (NICUs). The mean total log counts of organisms were 5.73 and 5.24 for the homemakers [defined as the person (usually the mother) who is the primary person responsible for arranging childcare, cooking, cleaning etc] and nurse hands, respectively (P<0.0001). Significantly more homemakers had Acinetobacter lwoffii, Enterobacter cloacae, Klebsiella pneumoniae, Pseudomonas aeruginosa, P. fluorescens/putida, and Staphylococcus aureus on their hands compared with the nurses (all P<0.05). However, significantly more nurses had Enterococcus faecalis, S. epidermidis, and S. warneri on their hands (P<0.05). Of note, the hands of nurses harboured significantly more S. epidermidis strains resistant to amoxicillin/clavulanate, cefazolin, clindamycin, erythromycin, and oxacillin and S. warneri resistant to amoxicillin/clavulanate, cefazolin, clindamycin, and oxacillin (P<0.05). Surprisingly, significantly more trimethoprim/sulfamethoxazole-resistant S. epidermidis and ciprofloxacin-resistant S. warneri was recovered from the hands of homemakers (P<0.05). This study demonstrates differences in prevalence, bacterial composition and antimicrobial resistance of hand flora of hospital personnel compared with homemakers. Moreover, the hands of homemakers may serve as community reservoirs for antimicrobial resistant strains of clinical importance.