To investigate the virulence genes, biochemical typing and multilocus sequence typing (MLST) distribution of Bacillus cereus from different sources in Wenzhou.Methods 127 strains of Bacillus cereus were identified and biochemical typed according to GB 4789.14-2014. Ten virulence genes were detected with polymerase chain reaction (PCR). Genetic typing was conducted with MLST.Results Of 127 isolates, NheA, NheB and NheC genes were accounted for 94.5% (120/127) and hblA, hblC and hblD genes were accounted for 9.4% (12/127). The emetic toxin gene was detected in 10 isolates (7.9%). Except 10 isolates which could not be typed, the rest 117 isolates were classified into type 2 (0.8%), type 5 (3.9%), type 8 (1.6%), type 9 (63.8%) and type 10 (22.0%) by biochemical typing. Seventy-two isolates could be classified into 28 sequence types (STs). ST26 (18.1%), ST144 (15.3%), ST92 (6.9%) and ST164 (6.9%) were the common STs. Cluster analysis on 28 STs generated four MLST groups and 13 singletons.Conclusion The positive rate of virulence genes was high in Bacillus cereus isolated from different sources in Wenzhou, and the genetic relationship was diverse.