[High specific PCR identification of Bungarus multicinctus and its adulterants].
OBJECTIVE
To develop a convenient and effective method for the identification of Bungarus multicinctus.


METHODS
Based on the sequence of Cyt b gene fragment of B. multicinctus and its adulterants, a pair of highly specific primer (HJL- and HJH-) were designed for distinguishing B. ulticinctus from other species of snake. To establish specific PCR reaction condition, the primers were employed to amplify the DNA templates extracted from B. multicinctus and 6 other species of snake, under different annealing temperature. Using this method, B. multicinctus was identified from 18 samples bought from many drugstores.


RESULTS
A 230 bp DNA fragment was amplified from B. multicinctus in PCR with annealed temperature at 67 degrees C, whereas no DNA fragment was amplified from other snake samples under the same reaction condition, B. multicinctus could be clearly distinguished from others by PCR reaction with the highly specific primers. In the present study, 18 sample, bought from different drugstores, were also identified by the highly specific PCR with the primers. The results indicated that 14 samples were B. multicinctus and the other 4 were adulterant, which was consistent with the conclusion of authentication based on morphological.


CONCLUSIONS
The primers designed in the present study were highly specific for B. multicinctus.
Feng
Cheng-qiang
CQ
Institute of Chinese Materia Medica, China Academy of Chinese Medicial Sciences, Beijing.
Tang
Xiao-jing
XJ
Huang
Lu-qi
LQ
Qian
Zhong-zhi
ZZ
Zhang
Ji
J
Cui
Guang-hong
GH
D000818:Animals; D001483:Base Sequence; D017838:Bungarus / Q000145:classification / Q000235:genetics*; D045303:Cytochromes b / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D017931:DNA Primers; D004340:Drug Contamination; D008419:Materia Medica; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D012911:Snakes / Q000145:classification / Q000235:genetics; D013045:Species Specificity
D017931:DNA Primers; D008419:Materia Medica; D004247:DNA; D045303:Cytochromes b