基于PCR指纹技术鹿鞭鉴定方法的建立及检测试剂盒的研制  

Establishment of Penis Cervi identification method based on PCR fingerprint technology and development of detection kit

作　　者：王雪松[1] 艾金霞[1] 薛晗 孙丽媛[1] 王艳双[1] 周亭亭[1] 李明成[1] 张丽华 WANG Xuesong;AI Jinxia;XUE Han;SUN Liyuan;WANG Yanshuang;ZHOU Tingting;LI Mingcheng;ZHANG Lihua(Department of Clinical Immunological Examination,Beihua University,Jilin City,Jilin Province,132013;Jilin Leining Food and Drug Testing Technology Services Co.Ltd,Jilin City,Jilin Province,132013,China)

机构地区：[1]北华大学临床免疫学检验教研室,吉林吉林132013 [2]吉林雷宁食品药品检测技术服务有限公司,吉林吉林132013

出　　处：《吉林医药学院学报》2021年第1期1-5,共5页Journal of Jilin Medical University

基　　金：吉林省中医药科技项目(2019132);吉林省教育厅“十三五”产业化项目(JJKH20180377KJ);吉林省科技厅发展计划项目(20190301014NY,20200404152YY,20200403047SF);省级大学生创新创业训练计划项目(201910201078).

摘　　要：目的建立鹿鞭及其伪品指纹鉴定方法,开发鹿鞭DNA检测试剂盒。方法采用改良的SDS方法从鹿鞭及牛鞭基因组中提取DNA。以鹿鞭线粒体细胞色素b作为靶基因,设计鹿鞭特异性引物,利用现代DNA指纹技术,研制了鹿鞭DNA提取和检测试剂,建立鹿鞭指纹鉴定方法,开发出鹿鞭DNA检测试剂盒。结果利用PCR指纹技术提取的鹿鞭基因组DNA为21000 bp,纯度为1.80±0.10之间;应用特异性引物可准确鉴别鹿鞭及牛鞭样品,鹿鞭特异性扩增产物DNA分子片段为307 bp,而牛鞭及阴性对照无扩增条带;自主研发鹿鞭DNA检测试剂盒经反复冻融依然有效,多次进行特异性、稳定性和重现性试验,结果完全一致。结论鹿鞭DNA检测试剂盒可作为鹿鞭鉴定的有效方法,该方法客观、准确、方便,可有效区分鹿鞭及其伪品。Objective To establish the fingerprint identification method of Penis Cervi and its counterfeit,and develop the detection kit of Penis Cervi DNA.Methods DNA was extracted from the genome of deer and bullwhip by improved SDS method.Using mitochondrial cytochrome b as the target gene,the specific primers were designed,and the DNA extraction and detection reagents were developed by using modern DNA fingerprinting technology.The method of deer whip fingerprint identification was established and the detection kit was developed.Results The genomic DNA extracted by PCR fingerprinting was 21000 bp with a purity of 1.80±0.10.The specific primers could be used to identify the samples of Penis Cervi and bullwhip,and the DNA fragment of the specific amplified product was 307 bp.There were no amplified bands in bullwhip and negative control.The self-developed Penis Cervi DNA detection kit was still effective after repeated freezing and thawing,and the results of specificity,stability and reproducibility tests were completely consistent.Conclusion The kit can be used as an effective method for the identification of Penis Cervi.The method is objective,accurate and convenient,and can effectively distinguish the Penis Cervi and its counterfeit.

关 键 词：鹿鞭 牛鞭 细胞色素B DNA检测试剂盒 PCR指纹技术 

分 类 号：R282.5[医药卫生—中药学]