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为建立金钱白花蛇品种的ＤＮＡ分子标记鉴定方法，本文通过提取金钱白花蛇及其伪品药材和原动物样品的模板ＤＮＡ，用通用引物扩增样品的Ｃｙｔ ｂ基因片断。 ＰＣＲ扩增所得产物纯化后直接测序，所得序列经对位排列和比较金钱白花蛇及其伪品的ＤＮＡ序列，发现Ｃｙｔ ｂ基因片断的种间差异显著大于种内个体间变异，因此该基因片断是蛇类药材鉴定中一个很好的分子标记。基于所得ＤＮＡ序列数据，设计了一对高度特异性的鉴别引物用于金钱白花蛇的ＰＣＲ鉴定。用该对引物对金钱白花蛇进行ＰＣＲ鉴定，在６０℃～６５℃的复性温度条件下，样品的误检和漏检率为０，能１００％地正确区分出正品与伪品药材，此外该方法还能检测出混合粉末中是否含有正品金钱白花蛇成分。研究结果表明用本鉴定引物对金钱白花蛇的ＰＣＲ鉴定方法简便、有效，实用性强，该方法还有可能成为中成药复方组分鉴别的一种新手段。
In order to establish the DNA molecular marker identification method for the species of white snakes, this article extracted the template DNA of snake and its fake medicinal materials and original animal samples, and amplified the Cyt b gene fragments of the samples with universal primers. The products amplified by PCR were directly sequenced after purification. The sequences obtained were aligned and compared with DNA sequences of P. alata and their counterfeits. It was found that the inter-species variation of the Cyt b gene fragment was significantly greater than the inter-individual variation within the species, and therefore the gene fragment was It is a good molecular marker in the identification of snake medicines. Based on the obtained DNA sequence data, a pair of highly specific identification primers was designed for the PCR identification of the snake. The PCR was used to identify the snakes of P. albinos using the pair of primers. Under the renaturation temperature of 60°C to 65°C, the false detection and missed detection rate of the samples was 0, and the genuine and fake medicinal materials could be correctly distinguished by 100%. The method can also detect whether the mixed powder contains a genuine money white snake ingredient. The results of the study showed that the PCR method used for identification of P. anatipestris with this identification primer was simple, effective, and practical. This method may also be a new method for identification of Chinese herbal medicine compound components.