We analyzed surveillance cultures for vancomycin-resistant enterococci (VRE) and methicillin-resistant Staphylococcus aureus (MRSA) collected during a multicenter trial to determine if three negative cultures collected at weekly intervals would predict clearance of VRE or MRSA from colonized patients. Seventy-two percent of VRE-colonized patients and 94% of MRSA-colonized patients were culture negative after three consecutive negative cultures.Infections with oxacillin (methicillin)-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) have increased dramatically in recent years, and together, they cause 12% of all health care-associated infections (2,8,16). Patients infected with MRSA or VRE, as well as asymptomatically colonized patients, serve as a reservoir for transmission of these bacteria to other hospitalized patients. To prevent spread, national guidelines recommend that health care providers use contact precautions during care of colonized and infected patients until it can be demonstrated that they are no longer colonized (5,10,14). Colonization with VRE can be prolonged (1, 3, 7), so the current recommendation by the Hospital Infection Control Practices Advisory Committee (HICPAC) is that isolation precautions should be maintained until VRE-negative results are documented with at least three consecutive negative cultures collected a minimum of 1 week apart (6,14,15). Persistent carriage with MRSA is also well documented (11, 13). Criteria for documenting clearance of MRSA from colonized patients are not established, although the standard of three negative weekly cultures is commonly applied to this population.The National Institute of Allergy and Infectious Diseases (NIAID) supported a large, cluster-randomized trial assessing strategies to reduce transmission of VRE and MRSA in 19 intensive care units (ICUs) (W. C. Huskins, C. M. Huckabee, N. P. O'Grady, P. R. Murray, H. Kopetskie, L. Zimmer, M. E. Walker, R. L. Sinkowitz-Cochran, J. A. Jernigan, M. Samore, D. Wallace, and D. A. Goldmann, submitted for publication). Stool or perianal swabs for VRE and anterior nasal swabs for MRSA were collected at all ICU sites from patients upon admission, weekly thereafter, and on discharge and were processed at a central laboratory. More than 22,000 swabs from approximately 9,900 patients were collected for VRE culture, and a similar number was collected for MRSA, during the 18-month study period. The trial did not include any systematic effort to "decolonize" patients, and topical antimicrobial agents (e.g., mupirocin, chlorhexidine, and vancomycin) were used infrequently in the participating ICUs. The systematic collection of specimens from ICU populations at multiple geographic sites presented an opportunity to evaluate the HICPAC recommendations for VRE and their applicability for MRSA colonization.Stool or perianal swabs for VRE were inoculated into bileesculin azide broth supplemented with 8 g/ml vancomycin and incubated at 35°C for 18 to 24 h. Broths were subcultured onto bile-...