Quality Evaluation of Galli Gigerii Endothelium Corneum by Allele-specific PCR Method / 中国实验方剂学杂志

ABSTRACT

Objective: To screen the specific reverse primers of Galli Gigerii Endothelium Corneum,duck gizzard membrane and goose gizzard membrane,and establish a specific PCR for molecular identifying Galli Gigerii Endothelium Corneum and its common adulterants. Method: Based on the mutation sites on the 12S rRNA sequence,specific polymerase Chain reaction(PCR) identify primers were designed for chicken,duck and goose gizzard membrane. The specific PCR reaction conditions were optimized,and the PCR identification method was explored and verified in terms of tolerance and feasibility. Thirty batches of Galli Gigerii Endothelium Corneum decoction pieces extracted from the test were identified. Result: Thirty batches of Galli Gigerii Endothelium Corneum decoction pieces were detected using chicken-specific primers, 273 bp of specific bands was amplified and visualized on the agarose electrophoregram. When duck and goose primers were used,no corresponding amplified band was detected. Conclusion: The allele-specific PCR method can be used as a rapid and accurate method to identify Galli Gigerii Endothelium Corneum. It is a promise method for special sampling tasks of Chinese herbal medicine and decoction tablets nationwide.