药用蛇及其水提物的DNA微型条形码鉴别研究

Identification of Medicinal Snakes and Water Extracts Based on DNA Mini-Barcode

目的:建立药用蛇及其水提物的DNA微型条形码鉴别方法。方法:通过比对药用蛇及常见11种混伪品的16S rRNA序列,建立约220 bp的蛇类DNA微型条形码,并与16S rRNA标准条形码比较,对药用蛇原料及水提物进行测序鉴定。采用MEGA-X软件分析遗传距离并以邻接法(NJ)构建系统发育树。结果:DNA微型条形码引物对药用蛇原料及水提物的扩增率为100%,鉴定率为100%。药用蛇及其混伪品的种内平均遗传距离均小于种间平均遗传距离,且各物种在NJ系统发育树中具有良好的单系性,并与16S rRNA标准条形码原料的鉴定结果一致。结论:该研究建立的蛇类DNA微型条形码可有效实现药用蛇原料及水提物等制品的物种鉴别,为保障蛇类原料、临床汤剂及配方颗粒等药品的安全性和有效性提供了技术支撑。

Objective:To establish a method for identification of medicinal snake and its water extract by DNA mini-barcode.Methods:By comparing the 16S rRNA sequences of medicinal snakes and 11 common adulterants,a mini-barcode of about 220 bp of snake DNA was established,and compared with the 16S rRNA standard barcode,the raw materials and water extracts of medicinal snakes were sequenced and identified.MEGA-X software was used to analyze genetic distance and construct phylogenetic tree by adjacency method(NJ).Results:The amplification rate and identification rate of DNA mini-barcode primers for medicinal snake raw materials and water extracts were 100%.The average intraspecific genetic distances of medicinal snakes and the adulterants were less than the average interspecific genetic distances,All species had good monophyletic characteristics in the NJ phylogenetic tree,which was consistent with the results of 16S rRNA standard barcode raw material identification.Conclusion:The snake DNA mini-barcode established in this study can effectively realize the species identification of medicinal snake raw materials and water extracts,and provides technical support for ensuring the safety and effectiveness of snake raw materials,clinical decoction and formula granules.