Development and evaluation of a DNA detection Kit on Identification of Agkistrodon Authenticity Based on Bioinformatics

摘要

We designed and evaluated a kind of Agkistrodon DNA detection kit. Bioinformatics was used to investigate the characteristics of cytochrome b (cyt b) gene in Agkistrodon. A series of primers were designed, and sequencing and blasting in conjunction with PCR technology were applied. Its' indexes including specificity, sensitivity and repeatability were evaluated according to the method recorded in Chinese Pharmacopoeia (2010 editions). The kit assay proved effective after repeated freezing and thawing 20 times, and repeatablity test indicated the same data through three times. The lowest amount was 0.025g of each tested specimen. The specificity test confirmed that 14 (77.8%) were real, and 4 was counterfeits in all samples. The developed DNA kit was accurate, effective in identification of Agkistrodon, compared with the Pharmacopoeia method.