Identification of Ophiophagus Hannah Cantor by Allele-specific PCR with Molecular Methods

OBJECTIVE To establish a specific polymerase chain reaction idetification of Ophiophagus hannah Cantor.

METHODS　 A total of 20 samples were collected, including Ophiophagus hannah Cantor, Naja naja, Agkistrodon acutus, etc. By comparing the cytochrome oxidaseⅠsequences of 20 snake samples, the specific single nucleotide polymorphisms of Ophiophagus hannah Cantor were searched and specific primers were designed; the key factors such as the number of cycles, annealing temperature were investigated to establish the optimal reaction system. The method was applied to identify the primordial of the medicinal powder from Ophiophagus hannah Cantor.

RESULTS When the annealing temperature was 58 ℃ and cycles number was 30, a specific band of 200 bp was amplified in the Ophiophagus hannah Cantor with specific primers, while other snake samples had no specific band.

CONCLUSION This method can distinguish the Ophiophagus hannah Cantor from other snake samples, and identify the primordial of the Ophiophagus hannah Cantor powder. It provides a scientific basis for the formulation and improvement of the quality evaluation system of Ophiophagus hannah Cantor medicinal materials and its decoction piece.