Abstract: According to the sequence of Cytochrome b in puffer fish published in GenBank, a puffer fish-specific primer pair of HT1-F and HT1-R was designed using Primer Premier 5.00 version for establishing a DNA based method to identify puffer fish species. Partial fragments of Cytochrome b gene in puffer fish samples including 9 major species from 3 genera and 2 unknown puffer fish samples harvested from Fujian province were amplified by PCR. The PCR products confirmed by agarose electrophoresis were used for sequencing. The length of the obtained DNA fragments was 423 bp in all samples. The homology of DNA sequences among these samples was analyzed using DNA MAN software, and a phylogenetic tree among these samples was also established. Eleven samples were divided into 3 groups, 89% of homology rate was observed between groups I and II; 85% of homology rate was observed between two groups and group III. In addition, 2 unknown puffer fish samples were probably from Gastrophysus or Takifugu genus. Collectively, these results provide a reference for the application of DNA sequencing technology to identify puffer fish components and its species present in processed food stuff.

Key words: puffer fish, PCR detection, DNA sequencing, species identification