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. 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.

中文翻译：

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ATPase 8基因引物电泳检测生化药物种源的特异性PCR方法

描述了一种 PCR 方法来识别各种动物和人体组织来源的生化药物的物种来源。本研究以精子片、复方牛胚肝提取物片、脾氨肽溶液、胎盘多肽注射液等四种商品化药物作为原理验证。引物旨在分别从牛肉、猪肉、羊肉和人类 DNA 中扩增线粒体细胞色素 b 和 ATPase 8 基因的保守区域。在给定的实验条件下，发现 ATPase 8 基因引物的特异性高于细胞色素 b 的特异性。对于猪肉、牛肉、羊肉和人体组织，ATPase 8 的扩增子大小分别为 212、271、293​​ 和 405 bp。用于物种鉴定的 DNA 样品的最小可检测浓度为 0.05–0.5 pg·μL-1。通过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。可以想象，这种用于肉类认证的 PCR 方法可以扩展到其他生化药物和原材料的质量控制。图形摘要开发了一种特异性 PCR 方法，用于通过靶向线粒体 ATPase 8 基因的物种特异性引物检测生化药物中的物种来源。PCR 产物通过凝胶电泳分离，物种来源通过与参考文献的比较来指示。开发了一种特定的 PCR 方法，用于通过靶向线粒体 ATPase 8 基因的物种特异性引物检测生化药物中的物种来源。PCR 产物通过凝胶电泳分离，物种来源通过与参考文献的比较来指示。