| 引用本文:杨重晖1，尹程程1，宋文博2，荆礼3，陈昊媛1，杨洋1，李芳宇2，刘莉2，赵大庆1，齐滨2.基于细胞色素C氧化酶亚基Ⅰ序列位点特异性聚合酶链反应鉴别鹿茸及其伪品[J].世界中医药,2020,(10):. |
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| 基于细胞色素C氧化酶亚基Ⅰ序列位点特异性聚合酶链反应鉴别鹿茸及其伪品 |
| Identification of Velvet Antler and Its Counterfeit Based on Site-specific PCR of COI Sequence |
| 投稿时间：2019-05-17 |
| DOI：10.3969/j.issn.1673-7202.2020.10.002 |
| 中文关键词: 鹿茸 细胞色素C氧化酶亚基Ⅰ DNA 位点特异性 聚合酶链反应 分子鉴定 |
| English Keywords:Cornu Cervi Pantotrichum COⅠ DNA Site-specific PCR Polymerase chain reaction Molecular identification |
| 基金项目:“十三五”国家重点研发计划项目（2018YFC1706604-05）；国家中药标准化项目（ZYBZH-Y-JL-26） |
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| 中文摘要: |
| 目的：基于细胞色素C氧化酶亚基Ⅰ(COⅠ)序列建立一种新的PCR鉴定方法，以快速鉴别出梅花鹿、马鹿茸片与驯鹿茸片。方法:采集不同来源的鹿茸片共60批。通过比较梅花鹿、马鹿与驯鹿等其他伪品的COⅠ基因序列差异，根据SNP鉴别位点设计梅花鹿茸、马鹿茸同驯鹿茸的鉴别引物Cne-F、Rt-F、Co-R，并对影响聚合酶链反应（PCR）结果的主要因素变性时间、退火温度、退火时间、延伸时间等进行方法学考察和优化。结果:该实验构建的双位点特异PCR鉴别体系，基于COⅠ的鉴别位点，可通过PCR扩增获得294 bp的梅花鹿、马鹿特异片段，而产生514 bp的驯鹿特异片段，伪品及阴性对照无条带。结论:该实验建立对梅花鹿、马鹿及驯鹿等其他伪品鹿茸的PCR鉴别方法简便、可靠。 |
| English Summary: |
| To establish a new PCR identification method based on COI sequence to quickly identify sika deer，Malurong Tablets and Xunlurong Tables.Methods:A total of 60 batches of Lurong Tablets from different sources were collected.By comparing the difference of COI gene sequences between fakes such as sika deer，red deer and reindeer，the identification primers Cne-F，Rt-F and Co-R of sika deer，red deer and reindeer velvet antler were designed according to the SNP identification location site.The main factors of polymerase chain reaction (PCR) results，such as denaturation time，annealing temperature，annealing time，and extension time，were investigated and optimized.Results:The two-site specific PCR identification system constructed in this study，based on the COI identification site，can obtain 294 bp sika deer and red deer specific fragments by PCR amplification，and produce 514 bp reindeer deer specific fragments，New Zealand red deer and other pseudo-pars and negative and control strip-free band.Conclusion:This experiment established a simple and reliable PCR identification method for other fake Cornu Cervi Pantotrichum such as sika deer，red deer and reindeer. |
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