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目的建立一种快速、高效的鉴别金钱白花蛇(银环蛇)与三种常见混伪品的多重PCR方法。方法采用动物DNA提取试剂盒提取金钱白花蛇、赤链蛇、赤链华游蛇与金环蛇DNA;根据各样品COI(动物线粒体细胞色素b基因)序列设计4对不同长度的特异性引物,建立并优化多重PCR反应体系,通过电泳检测扩增产物分子量差异实现4种蛇类的快速鉴别。结果针对金钱白花蛇、赤链蛇、赤链华游蛇与金环蛇所设计的4对引物分别扩增出136、195、244和315bp的目的条带;每对引物特异性扩增,灵敏度为皮克级。结论首次建立了金钱白花蛇与常见三种相似混伪品的多重PCR方法。此方法特异性高,为多重PCR技术体系在中药材金钱白花蛇的快速检测提供参考。
OBJECTIVE: To establish a rapid and efficient multiplex PCR method for identifying Meriones niloticus (Bungarus) and three common mixed fake products. Methods The genomic DNA was extracted from the genus Ardisia serrata, Ardisia serici, Aristochina nakai and Aristolochia sericata by animal DNA extraction kit. Four pairs of specific primers with different length were designed according to COI (animal mitochondrial cytochrome b gene) The multiplex PCR system was established and optimized, and the difference of the molecular weight of the amplified product was detected by electrophoresis to rapidly identify four species of snakes. Results The target bands of 136, 195, 244 and 315 bp were amplified respectively from 4 pairs of primers designed for the Chinese white Huacha, red chain snake, red chain snake and jinx snake; each pair of primers specific amplification, sensitivity Pico level. Conclusion The multiplex PCR method was established for the first time in the world. The method is highly specific and provides a reference for the rapid detection of the money Chinese white snake in the multi-PCR system.