ABSTRACT
OBJECTIVE:To provide new identification method for processed medicinal material Chinese polyphaga(Eupolyph-aga sinensis,Steleophaga plancyi) and their adulterants by establishing molecular identification method based on Cytb genes. METHODS:The total DNA of Chinese polyphaga and their adulterants was extracted using modified saturation sodium chloride method. The Cytb genes of all samples were amplified with PCR using general primers REVCB2H and REVCBJ. The phylogenetic tree of all samples was constructed with Neighbor-Joining(NJ)method using MEGA 5.1 software. The sequences of the Cytb gene of all sampled were compared by using DNAMAN sofetware. The difference between genuine product and their adulterants were analyzed,and the specific primers Esin-F and Esin-R were designed for molecular identification in different regions. RESULTS:DNA extracted from processed medicinal insects was successful to amplify Cytb gene segments. The phylogenetic tree of all sam-ples was consistent with their genetic relationship. A fragment was amplified only from genuine product but not from other adulter-ants with the designed specific primers Esin-F and Esin-R. CONCLUSIONS:DNA extraction method from processed Chinese polyphaga and their adulterants have been established. Designed specific primers are highly specific to genuine product Chinese polyphaga,and can be used for the identification of Chinese polyphaga and their adulterants.