Specific PCR method for detection of species origin in biochemical drugs via primers for the ATPase 8 gene by electrophoresis
Specific PCR method for detection of species origin in biochemical drugs via primers for the ATPase 8 gene by electrophoresis
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ATPase 8 基因引物电泳检测生化药物物种来源的特异性 PCR 方法
DOI：
10.1007/s00604-019-3738-5
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发表时间：
2019-09-01
影响因子：
5.7
通讯作者：
Bai, Liping
中科院分区：
文献类型：
--
作者：
Ai, Limei;Liu, Juanjuan;Bai, Liping
描述了一种用于鉴定各种动物和人类组织衍生生化药物的物种来源的聚合酶链式反应方法。本研究以四种商品化药物：片、复方胎牛肝提取物片、脾氨基肽溶液、胎盘多肽注射液作为原理验证。设计了从牛肉、猪肉、羊肉和人类DNA中分别扩增线粒体细胞色素b和ATPase 8基因保守区的引物。在给定的实验条件下，ATPase 8基因的特异性高于细胞色素b。ATPase 8在猪肉、牛肉、羊肉和人体组织中的扩增片段大小分别为212、271、293和405个碱基。物种鉴定的最低检出浓度为0.05pg·μL-1。这种方法可以在提取的模板DNA浓度极低的情况下区分物种来源。可以想见，这种用于肉类认证的PCR方法可以推广到其他生化药物和原料的质量控制。通过针对线粒体ATPase 8基因的物种特异性引物，建立了一种检测生化药物中物种来源的特异聚合酶链式反应方法。用凝胶电泳法对扩增产物进行了分离，并与文献进行了比较，指出了其物种来源。
A PCR method is described to identify the species origin of various animal and human tissue-derived biochemical drugs. Four commercialized drugs, including spermary tablets, compound embryonic bovine liver extract tablets, spleen aminopeptide solution, and placenta polypeptide injection, were used as a proof-of-principle in this study. Primers were designed to amplify conservative regions of mitochondrial cytochrome b and ATPase 8 genes from beef, pork, lamb and human DNA, respectively. The specificity of primers for ATPase 8 gene is found to be higher than those for cytochrome b under the given experimental conditions. The amplicon sizes of ATPase 8 were 212, 271, 293 and 405 bp for pork, beef, lamb and human tissue, respectively. The minimum detectable concentration of DNA sample for species identification is 0.05-0.5 pg·μL-1. The species origin can be distinguished by this method in extremely low concentrations of template DNAs extracted. Conceivably, this PCR method for meat authentication may be extended to quality control of other biochemical drugs and raw materials. Graphical abstract A specific PCR method was developed for the detection of species origin in biochemical drugs via species-specific primers targeting mitochondrial ATPase 8 genes. The PCR products were separated by gel electrophoresis and species origins were indicated by comparison to references.