To quantify the sensitivity of surveillance cultures for the detection of multidrug-resistant gram-negative (MDRGN) bacteria, perianal/rectal swabs were collected from patients with positive clinical cultures for MDRGN species. Surveillance cultures identified colonization with the same genetically related MDRGN species in 29 of 37 MDRGN clinical culture isolates (78%). There was a trend toward less antimicrobial exposure among patients with false-negative surveillance culture results (P聽= .06).