OBJECTIVE To develop a convenient and effective method for the identification of Zaocys dhumnades(black snake).METHODS Based on the sequence of mitochondrial 12S rRNA gene fragment of Zaocys dhumnades and its adulterants,a pair of highly specific primer(HWL-1 and HWH-1) were designed for distinguishing Zaocys dhumnades from other species of snake.A specific PCR reaction condition was established.The primers were employed to amplify the DNA templates extracted from Zaocys dhumnades and 10 other species of snake,under different annealing temperatures.Using this method,Zaocys dhumnades from thirteen samples bought from drugstores was identified.RESULTS About 320 bp DNA fragment was amplified from Zaocys dhumnades in PCR with annealed temperature at 65 ℃,whereas no any DNA fragment was amplified from other snake samples under the same reaction condition.Zaocys dhumnades was clearly distinguished from other by PCR reaction with the highly specific primers.In the present study,thirteen samples,bought from different drugstores,were also identified by the highly specific PCR with the primers.The results indicated that 9 samples were Zaocys dhumnades and the others were adulterants,which was consistent with the conlusion of authentication based on morphology.CONCLUSION The primers designed in the present study are highly specific for Zaocys dhumnades.This method is simple,fast and reproducable.