ObjectivesTo examine factors associated with isolation precaution use in nursing home (NH) residents with multidrug-resistant organism (MDRO) infection. DesignRetrospective, cross-sectional analysis. SettingNursing homes with Centers for Medicare and Medicaid Services' certification from October 2010 to December 2013. ParticipantsElderly, long-stay NH residents with positive MDRO infection assessments. MeasurementsData were obtained from the Minimum Data Set (MDS) 3.0, Certification and Survey Provider Enhanced Reporting, and Area Health Resource File. Multivariable regression with facility fixed effects was conducted. ResultsThe sample included 191,816 assessments of residents with MDRO infection, of which isolation use was recorded in 12.8%. Of the NHs reporting MDRO infection in the past year, 31% used isolation at least once among residents with MDRO infection. Resident characteristics positively associated with isolation use included locomotion (23.6%, P < .001) and eating (17.9%, P < .001) support. Isolation use was 14.3% lower in those with MDRO history (P < .001). Residents in NHs that had received an infection control-related citation in the past year had a greater probability of isolation use (3.4%, P = .02); those in NHs that had received a quality-of-care citation had lower probability of isolation use (-3.3%, P = .03). ConclusionThis is the first study to examine the new MDS 3.0 isolation and MDRO items. Isolation was infrequently used, and the proportion of isolated MDRO infections varied between facilities. Inspection citations were related to isolation use in the following year. Further research is needed to determine whether and when isolation should be used to best decrease risk of MDRO transmission and improve quality of care.