To establish a new method for geneotyping of Vibrio vulnificus.Methods PCR-denaturing gradient gel electrophoresis(PCR-DGGE)was used to elucidate the molecular characteristics of the polymorphism of 16S-23S rDNA intergenic spacer regions(ISR)from 18Vibrio vulnificus strains. Meanwhile, detection of virulence factors and antimicrobial susceptibility test of isolates were conducted to verify the relationship of the strains.Results The Vibrio vulnificus strains could be amplified into 4different bands by ISR-PCR, including 900,0, 650and 550bp. At the same time, Vibrio vulnificus ISR-DGGE sequences showed significant differences between the strains, and all 18strains were typed into 16types by DGGE. Clustering analysis divided them into two categories with similarity of 0.65and 0.71. The results of antimicrobial susceptibility test implied that drug resistant types of sj6, sj11and sj12were different from the other 15strains, which was consistant to the ISR-DGGE.ConclusionThis molecular technique could be applied to investigation and identification of Vibrio vulnificus, and provide a new method for geneotyping.