蕲蛇实时荧光定量PCR快速鉴别方法的建立  

Establishment of a Method of Rapid Identification of Agkistrodonby Real-Time Fluorescence Quantitative PCR

作　　者：裴社强 杨宝 PEI Sheqiang;YANG Bao(Shanxi Inspection and Testing Center institute of Drug InspectionTechnology,Taiyuan Shanxi,030000,China)

机构地区：[1]山西省检验检测中心药品检验技术研究所,山西太原030000

出　　处：《质量安全与检验检测》2022年第3期11-14,30,共5页QUALITY SAFETY INSPECTION AND TESTING

摘　　要：为建立中药材蕲蛇的快速鉴别方法,本文基于CO1片段序列特征,采用Oligov 7.0.1软件设计了蕲蛇的特异性引物和TaqMan探针,用以建立TaqMan探针荧光PCR检测蕲蛇成分的方法。结果显示,引物和探针对蕲蛇检测的特异性强,与其他近源性蛇类DNA无非目标扩增,与其他PCR方法相比,所用设备少、污染小,能够在20~40 min得出检测结果,且检测结果50 ng稀释105倍后仍可检测出蕲蛇成分,检测限达到0.5 pg/反应,可见该方法具有准确、快捷、灵敏的特点,可应用于蕲蛇检测,对于保证蕲蛇药材的质量及用药安全具有重要意义。Based on the sequence characteristics of CO1 fragments,agkistrodon-specific primers and TaqMan probes were designed using Snap gene software Oligov 7.0.1.And then the rapid identification method of agkistrodon was established by real-time fluorescence quantitative PCR.The results showed that the primers and probes had strong specificity for the detection of agkistrodon.The results showed that the primers and probes had high specificity for agkistrodon without non-targeted amplification for other proximal snakes DNA.Compared with other PCR methods,this method required less equipment,caused less pollution,and it only took 20-40 minutes to get the results.The method is highly sensitive,the lower limit of detection was 0.5 pg Per reaction.The method is accurate,fast,sensitive,therefore it can be used in the detection of agkistrodon.It is important to ensure the quality and safety of agkistrodon.

关 键 词：蕲蛇 实时荧光定量PCR 快速鉴别 

分 类 号：R927.11[医药卫生—药学]