To investigate Clostridium ­difficile (C. difficile)­associated diarrhea (CDAD) in human immunodeficiency virus (HIV)­infected inpatients in Shanghai, C. difficile and general intestinal tract pathogenic bacteria were isolated by selective medium from abnormal fecal specimens collected from 250 HIV ­infected inpatients from March 2012 to March 2013. The susceptibilities of C. difficile to metronidazole, vancomycin, clindamycin and moxifloxacin were determined by E­-test method. Molecular typing was performed by multi­locus sequence typing (MLST). The genes tcdA and tcdB of toxins A and B were detected by conventional polymerase chain reaction (PCR). 29 clinical strains of C. difficile were isolated from 250 fecal specimens (11.6%), in which 17 strains were tcdA^＋/tcdB^＋ and 12 strains were tcdA^－/tcdB^－. All strains were susceptible to metronidazole and vancomycin. Resistance to clindamycin and moxifloxacin was found in 69.0% and 31.0% strains respectively. A total of 12 different sequence types (ST­2, ­3, ­7, ­8, ­26, ­35, ­37, ­38, ­39, ­54, ­81, and ­124) were identified. The most prevalent ST types were ST­54 (6/29, 20.7%) and ST­39 (5/29, 17.2%). The results suggest that the rate of positive C. difficile detection is high and should be monitored strictly to prevent outbreak.