Abstract: Objective To characterize O3∶K6 serotype of V. parahaemolyticus isolated from cases of food poisoning during 2006-2012 in Nanshan District, Shenzhen. Methods Strains of V. parahaemolyticus serotype O3∶K6 were collected. Multi-PCR, pulsed field gel electrophoresis (PFGE), and Kirby-Bauer were performed. Results A total of 44 strains of V. parahaemolyticus (O3∶K6) obtained from diarrhea patients with food poisoning were collected. All strains were tdh gene positive and trh gene negative. PFGE indicated that the 44 strains could be divided into 2 clone complexes. Clone 1 (C1) had 19 strains, with 17-20 bands for each strain, while clone 2 (C2) had 25 strains, with 17-18 bands for each one. Differences between C1 and C2 were found in position of 200, 190, and 100 Kb, while the other bands were similar. All the 44 strains (100.00%) were sensitive to cefrazidime, cefepime，gentamicin，levoflocacin，tetracycline，chloramphenicol, and sulphamethoxazole, 72.73% (32/44), 18.18% (8/44), and 18.18% (8/44) of strains were resistant to ampicillin, kanamycin, and streptomycin, and 9.09% (4/44), 65.91% (29/44), and 65.91% (29/44) of strains were moderately sensitive to the three antibiotics, respectively. Conclusion The main PFGE clone complexes of strains of V.parahaemolyticus serotype O3∶K6 isolated in the local area were C1 and C2, with tdh gene positive and trh gene negative.