Abstract:
DNAs extracted from both “Jinqian Baihuashe” (
Bungarus parvus) and its adulterants and original animals of the crude snake drugs were used as templates for Cyt
b gene fragment amplification. The sequence data of the fragments showed that the differences of the sequence between
Bungarus parvus and its adulterants were far greater than that between intraspecific variations of
Bungarus parvus. Therefore, the Cyt
b gene fragment was a good molecular genetic marker for the authentication of
Bungarus parvus. On the basis of the sequence data, a pair of specialized primers, BuL-1 and BuH-1 was designed for the PCR identification of
Bungarus parvus. The effectiveness of the primers were examined at a series of anneal temperatures. The results showed that
Bungarus parvus samples could be absolutely distinguished when the anneal temperatures were 60℃~65℃, whereas no incorrect or missing discrimination was found at these temperatures. The results also showed that the powder of
Bungarus parvus which was mixed with powders of three other crude snake drugs may be detected by the PCR identification. This indicates that PCR identification may be a new method for examining the compositions of Chinese patent medicine.