Because the epidemiology of Clostridium difficile infection (CDI) is region-specific, the present study was undertaken to examine the epidemiology of C difficile outbreaks in Beijing, China. Eighty nonduplicate isolates were collected from March, 2016 to December, 2016. The molecular type and phylogenetic analysis were evaluated by multilocus sequence typing (MLST). The minimum inhibitory concentrations (MICs) for 11 antibiotics and the resistance mechanisms were investigated. Sixty-five toxigenic strains (81.25%), including 22 tcdA(-)B(+)CDT(-) strains (27.5%) and 43 tcdA(+)B(+)CDT(-) strains (53.75%), and also 15 nontoxigenic strains (tcdA-B-CDT-; 18.75%) were detected. MLST identified 21 different sequence types (STs), including 2 novel types (ST409 and ST416). All isolates were susceptible to metronidazole, vancomycin, fidaxomicin, piperacillin/tazobactam, and meropenem, and all were effectively inhibited by emodin (MICs 4-8mg/mL). The resistance rates to rifaximin, ceftriaxone, clindamycin, erythromycin, and ciprofloxacin were 8.75%, 51.25%, 96.25%, 81.25%, and 96.25%, respectively; 81.25% (65/80) of isolates were multidrug-resistant. Amino acid mutations in GyrA and/or GyrB conferred quinolone resistance. One novel amino acid substitution, F86Y in GyrA, was found in 1 CIP-intermediate strain. The erm(B) gene played a key role in mediating macrolide-lincosamide-streptogramin B (MLSB) resistance. Erm(G) was also found in erm(B)-negative strains that were resistant to both erythromycin and clindamycin. RpoB mutations were associated with rifampin resistance, and 2 new amino mutations were identified in 1 intermediate strain (E573A and E603N). Regional diversity and gene heterogeneity exist in both the ST type and resistant patterns of clinical C difficile isolates in Northern China.