Authentication of an animal crude drug, Zaocys, by diagnostic PCR

A pair of diagnostic primers for distinguishing the Chinese crude drug Zaoc ys (Zaocys dhumandes) from its substitutes was designed based on the sequen ce data of the original animal of the drug and substitutes. Total DNAs were extracted from genuine crude drug and 5 of its substitutes, as well as fro m 12 species of original animal of the snake crude drug. Diagnostic PCRs we re performed using the primers with these total DNAs as a template, anneali ng at 60-65 degrees C, Positive amplifications were obtained from all DNA t emplates of Zaocys, whereas negative amplifications were obtained from that of others, The results indicate that Zaocys samples could be definitely di stinguished from its substitutes by diagnostic PCR, and no incorrect discri mination was found under the same reaction conditions. The advantages of th e method in the authentication of crude drugs are also discussed in the pre sent paper.