Yanqing Chen, Bo Wang, Ping Wang, Bisheng Huang, Hegang Liu, Chao Xiong, Mingli Wu, Nili Wang, Di Liu, Zhigang Hu, Shilin Chen
a College of Pharmacy , Hubei University of Chinese Medicine , Wuhan , China.
b Hubei Institute for Drug Control , Wuhan , China.
Mitochondrial DNA A DNA Mapp Seq Anal. 2019 Mar;30(2):367-374. doi: 10.1080/24701394.2018.1532417. Epub 2019 Jan 27.
Effective identification methods for snake species are lacking, exacerbating the extermination of medicinal and commercially valuable snake species. Hence, it is imperative to find fast and reliable methods to distinguish snake samples available on the market. Seventy-three samples from four families belonging to 13 genera were collected in China and found to contain common medicinal snakes and their adulterants. Cytochrome oxidase I (COI) was utilized as a DNA barcode to analyse these common snakes, and a DNA mini-barcode was employed for fast detection. Then, the DNA mini-barcode assays were coupled with a high-resolution melting (HRM) analysis (Bar-HRM) to realize the rapid discrimination of these snake species. The results showed the power of DNA barcoding with COI, which was capable of distinguishing all collected snake samples, and the combined Bar-HRM method can successfully identify the adulterants and different snake species. In particular, Bar-HRM revealed Bungarus fasciatus adulterants in B. multicinctus at concentrations as low as 1.6%. Moreover, the results of the study confirmed the effectiveness of the technique in terms of the rapid identification of snakes, which has great potential for ensuring the safety of commercially valuable snake species.
目前缺乏有效的蛇类物种鉴定方法,这加剧了药用和商业价值高的蛇类物种的灭绝。因此,必须找到快速可靠的方法来区分市场上现有的蛇类样本。在中国收集了来自13个属4个科的73个样本,发现其中含有常见的药用蛇及其掺假品。利用细胞色素氧化酶I(COI)作为DNA条形码来分析这些常见蛇类,并采用DNA微型条形码进行快速检测。然后,将DNA微型条形码检测与高分辨率熔解(HRM)分析(Bar-HRM)相结合,以实现对这些蛇类物种的快速鉴别。结果显示了COI DNA条形码的鉴别能力,它能够区分所有收集到的蛇类样本,并且结合的Bar-HRM方法可以成功鉴定掺假品和不同的蛇类物种。特别是,Bar-HRM揭示了在多带腹链蛇中掺有白链蛇,其浓度低至1.6%。此外,该研究结果证实了该技术在快速鉴定蛇类方面的有效性,在确保商业价值高的蛇类物种安全方面具有巨大潜力。