快速鉴定鹿茸真伪及其种属检测试剂的试验研究  

Experimental study on rapid identification of antler authenticity and its species detection reagents

作　　者：徐宁 马玉贺 马秋贺 艾金霞 母润红[2] 高丽君 夏薇 XU Ning;MA Yu-he;MA Qiu-he;AI Jin-xia;MU Run-hong;GAO Li-jun;XIA Wei(School of Medical Technology,Beihua University,Jilin 132013,China;School of Basic Medicine,Beihua University,Jilin 132013,China)

机构地区：[1]北华大学医学技术学院,吉林132013 [2]北华大学基础医学院,吉林132013

出　　处：《药物分析杂志》2023年第8期1435-1445,共11页Chinese Journal of Pharmaceutical Analysis

基　　金：吉林省科技发展计划项目资助(20230401096YY、20200403047SF、20210204185YY);吉林省科技厅重点科技成果转化项目资助(20170307001YY)。

摘　　要：目的:建立1种双重PCR检测体系,根据琼脂糖凝胶电泳目的条带位置及数量快速鉴定花鹿茸、马鹿茸及常见伪品驯鹿茸。方法:采用碱变性法提取鹿茸样品基因组DNA,以鹿科动物Cytb和COI基因为靶序列,应用Primer 3.0,根据SNP位点设计特异性鉴别引物(Cy-1、COI-1),并摸索及优化PCR反应体系及条件,同时,采用克隆及测序验证检测结果的准确性。结果:建立的双重PCR扩增体系显示,花鹿茸在电泳图谱408 bp及146 bp处可见2条扩增条带,马鹿茸则在146 bp处见1条目的条带,而伪品鹿茸则无扩增条带,与质粒克隆测序验证的结果一致。且市售鹿茸样品的检测结果与鉴定结果相吻合。结论:本试验自主构建的双重PCR检测体系简便、迅速,检测试剂结果准确,稳定性良好,在分子水平实现一步法鉴定花鹿茸、马鹿茸及其常见伪品,具有较大实用价值。Objective:To establish a duplex PCR detection system for rapid identification of pilose antler,red deer antler and common counterfeit reindeer antler according to the location and number of target bands on agarose gel electrophoresis.Methods:The genomic DNA of velvet antler samples was extracted by alkali denaturation method.The Cytb and COI genes of deer animals were used as target sequences,and Premier 3.0 was used to de⁃sign specific identification primers(Cy-1,COI-1)based on SNP sites.The PCR reaction system and condi⁃tions were explored and optimized.At the same time,the accuracy of the test results was verified by cloning and sequencing.Results:The established duplex PCR amplification system showed that there were two amplified bands at 408 bp and 146 bp in the electrophoresis map of velvet antler,one band at 146 bp in velvet antler,and no amplified band in counterfeit velvet antler,which was consistent with the results of plasmid cloning and sequen⁃cing.The detection results of commercial antler samples were consistent with the identification results.Conclusion:The duplex PCR detection system constructed in this experiment is simple and rapid,and the detec⁃tion reagent results are accurate and stable.It is of great practical value to realize one-step identification of velvet antler,red deer antler and their common counterfeits at the molecular level.

关 键 词：花鹿茸 马鹿茸 双重聚合酶链式反应 位点特异性 检测试剂 

分 类 号：R917[医药卫生—药物分析学]