科研动态

为建立可以区分H7N9流感病毒强毒株和弱毒株的方法，通过比对GenBank和GISAID中H7N9流感病毒的HA基因序列以及本实验室保存的病毒序列，设计1对特异性引物和2条探针，建立了区分H7N9流感病毒强弱毒株的实时荧光RT-PCR方法，同时对该方法的反应体系和反应参数进行优化，并开展特异性和敏感性试验以及临床应用试验。特异性试验显示，该方法具有良好的特异性，对其他常见亚型的禽流感病毒和其他禽病病毒检测均为阴性。敏感性试验显示，该方法对H7N9流感强毒和弱毒HA基因的检测下限分别为0.004 fg 和0.1 fg RNA模板，高于两种商品化试剂盒的灵敏度。利用该方法对40株经实验室分离鉴定的H7N9流感病毒进行对比验证，发现检出率和符合率均为100%。结果表明，该方法具有特异、敏感、准确的优点，可用于H7N9流感病毒的快速检测，同时还可区分强弱毒株，对H7N9流感病毒，尤其是高致病性流感病毒的早期诊断和有效防控具有重要意义。

Development of a Real-time RT-PCR Method

for Identification of Virulent and Avirulent Strains of H7N9Influenza Virus

In order to develop a method for identifying virulent andavirulent strains of H7N9 influenza virus，a pair ofspecific primers and probes were designed by comparing the HA gene sequences ofH7N9 influenza virus in Genbank and GISAID and the sequence of which wasisolated and sequenced by our laboratory，a real-timeRT-PCR method was developed，meantime，the reaction system and parameters of the method were optimized，and the specificity and sensitivity as well as its clinicalapplication were tested. The specificity test showed that，the development method didn't react with other common subtypes ofavian influenza virus and other avian virus，showing agood specificity. Sensitivity test showed that，by theRT-PCR method，the lower detection limits of HA genes ofvirulent and attenuated H7N9 strains were 0.004 fg and 0.1 fg respectively，which were all lower than the detection limits of the two kinds ofcommercial kits. By comparing and verifying 40 strains of H7N9 influenza virusisolated and identified by the laboratory，it was foundthat both the detection rate and coincidence rate were 100%. Therefore, it wasconcluded that the method could be used for rapid detection of H7N9 virus andidentifying the virulent and avirulent strains due to its advantages of goodspecificity，sensitivity and high accuracy，which was of great significance for early diagnosis and effectiveprevention and control of H7N9 virus，especially thehighly pathogenic virus.

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