ABSTRACT: The polymerase chain reaction (PCR) and direct sequence analysis were used to determine the amount of genetic variation in a 376-nucleotide region of the mitochondrial cytochrome b gene in fresh, frozen and steam sterilized meats of puffer Takifugu rubripes. A diversity of sequence between fresh, frozen and steam sterilized meats was not found. The puffer specimens collected from either culture ponds in Taiwan or wild seawaters and culture waters in Japan had the same sequence of 376-nucleotide region. Restriction enzyme BstZI was found to cut the amplified region of cytochrome b gene in fresh, frozen and steam sterilized meats of puffer T. rubripes, while restriction enzyme AatII did not. This indicates that the sequence and restriction site analyses might be used to authenticate species of different processed meats of puffer T. rubripes.