H7N9亚型流感病毒荧光定量RT-PCR检测方法的建立  

Establishment of a Novel RT-PCR Assay for Detection of H7N9 Subtype Influenza Virus

作　　者：雷宇平[1] 图门巴雅尔 张仲萍[1] 王锦 宁艳[1] 雷冲[1] 胡明明 孙杰[1] 王治维[1] 赵凯 张昱 王仲兵[1] Lei Yuping;Tumen Bayaer;Zhang Zhongping;Wang Jin;Ning Yan;Lei Chong;Hu Mingming;Sun Jie;Wang Zhiwei;Zhao Kai;Zhang Yu;Wang Zhongbing(Shanxi Center for Animal Disease Prevention and Control,Taiyuan,Shanxi 030027,China)

机构地区：[1]山西省动物疫病预防控制中心,山西太原030027

出　　处：《中国动物检疫》2021年第8期90-94,共5页China Animal Health Inspection

基　　金：山西省重点研发计划项目(201803D221026-2)。

摘　　要：为建立一种快速检测H7N9亚型流感病毒的通用荧光RT-PCR方法,根据GenBank数据库中收录的相关基因组序列,应用Primer 5.0软件设计合成针对H7和N9序列的特异性引物和Taq Man探针,并对该方法开展特异性、敏感性、重复性试验。结果显示:H7和N9核酸片段最低检测限均为1.7×10^(3) copies/mL;使用该方法重复检测10次,H7和N9核酸片段检测的变异系数(CV)<1%,说明稳定性良好;利用第二代人感染H7N9流感病毒核酸检测试剂国家参考品样品,测得该方法灵敏度为100%(10/10)、特异度为100%(20/20)。结果表明,本研究建立的H7N9亚型流感病毒荧光定量RT-PCR方法快速、准确、稳定性高、特异性好,为控制流感病毒传播增添了技术手段。In order to establish a novel RT-PCR assay to rapidly detect H7N9 subtype influenza virus,specific primers and Taq Man probes for H7 and N9 sequences were designed and synthesized by Primer 5.0 according to relevant genome sequences collected in GenBank database,followed by the test of sensitivity,specificity and repeatability.The results showed that,for H7 and N9 nucleic acid fragments,the detection limits were both 1.7×10^(3) copies/mL;the coefficient of variation(CV)was less than 1%after detection for 10 times,which indicated that the method was with good stability;the sensitivity and specificity were 100%(10/10)and 100%(20/20),respectively as measured by the reagents from 2 nd National Reference Panel for Avian Influenza A(H7N9)Viral Nucleic Acids Detection Kit.In conclusion,the established method could be used as a technical tool to control any spreading of influenza virus due to its advantages of rapidity,accuracy,good stability and specificity.

关 键 词：H7N9亚型流感 荧光定量RT-PCR 敏感性 特异性 重复性 

分 类 号：S852.65[农业科学—基础兽医学]