金钱白花蛇、乌梢蛇、蕲蛇的三重PCR鉴定方法的建立  

Establishment of triple PCR method for identification of Bungarusparvus,Zaocys and Agkistrodon

作　　者：刘美琳 王艳双[1,3] 姜海瀛 孙丽媛 李明成 张丽华[4] LIU Mei-lin;WANG Yan-shuang;JIANG Hai-ying;SUN Li-yuan;LI Ming-cheng;ZHANG Li-hua(School of Medical Faculty,Beihua University,Jilin 132013,China;School of Laboratory Medicine,Beihua University,Jilin 132013,China;Chinese Medicine DNA Fingerprint Detection Technology Technology Innovation Center,Beihua University,Jilin 132013,China;Jilin Leining Food and Drug Testing Technology Service Co.,Ltd.,Jilin 132013,China)

机构地区：[1]北华大学基础医学院,吉林132013 [2]北华大学医学技术学院,吉林132013 [3]北华大学吉林省中药DNA指纹检测技术科技创新中心,吉林132013 [4]吉林雷博科技有限公司,吉林132013

出　　处：《药物分析杂志》2022年第5期795-801,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：吉林省科技发展计划项目(20190303093SF,20190301014NY,20200404152YY,20200403047SF);吉林省科技创新中心建设项目(20190902018TC)。

摘　　要：目的:建立金钱白花蛇、乌梢蛇、蕲蛇的三重聚合酶链式反应(PCR)鉴别方法。方法:以线粒体Cytb基因为靶基因,设计特异性引物,优化引物最适退火温度与最佳浓度比,并用该方法进行混合样品的检测。结果:金钱白花蛇、乌梢蛇与蕲蛇的引物(浓度均10μmol·L^(-1))在57℃时特异性均非常好,最佳浓度比为0.6∶1.0∶1.8时分别扩增出185、235、343 bp的特异性条带,混淆品及空白对照无条带。该方法能同时、准确地鉴定出混合样品的蛇源成分。结论:本方法可同时、准确、快速地鉴定金钱白花蛇、乌梢蛇和蕲蛇样品,并适合粉末样品鉴别。Objective:To establish a triplex polymerase chain reaction(PCR)method for the identification of Bungarusparvus,Zaocys and Agkistrodon.Methods:Taking mitochondrial Cytb gene as the target gene,specific primers were designed,and the optimal annealing temperature and concentration ratio of the primers were optimized.The mixed samples were detected by this method.Results:The specificity of the primers(10μmol·L^(-1))was good at 57℃.The specific bands at 185,235 and 343 bp were amplified at the optimal concentration ratio of 0.6∶1.0∶1.8,respectively.The adulterants and the blank control shoewed no bands.The method could simultaneously and accurately identify the snake-derived components in the mixed samples.Conclusion:The method can be used to identify the samples of Bungarusparvus,Zaocys and Agkistrodon simultaneously,accurately and rapidly,and is suitable for the identification of powder samples.

关 键 词：金钱白花蛇 乌梢蛇 蕲蛇 CYTB基因 三重聚合酶链式反应(PCR) 鉴定 

分 类 号：R917[医药卫生—药物分析学]