A serial prospective survey of nasal colonization of hospital personnel by methicillin-resistant coagulase-negative staphylococci (MRCNS) was conducted at a Veterans Affairs medical center on three occasions over a 16-month period. The epidemiological typing systems used to assess relatedness included antimicrobial susceptibility profiles; biotyping; phage typing; plasmid profiles; restriction fragment length polymorphism (RFLP) analysis with ribosomal RNA; and plasmid hybridization with a 1.68-MD plasmid as the DNA probe. Forty-three percent of all personnel and 62% of all nurses were colonized with MRCNS. Nurses on the wards (72%) and in the intensive care unit (73%) were significantly more likely to be colonized with MRCNS than nurses who had less contact with patients or those who worked in the operating room. The molecular epidemiological typing systems indicated some degree of relatedness among the strains. Specifically, riboprobe analysis revealed a Dice coefficient of > 90%. However, each typing system detected dissimilarity among strains. Further studies are needed to determine the role that such human reservoirs of MRCNS serve in horizontal transmission to and subsequent infection of hospitalized patients.