用两种聚合酶链反应方法检测流感病毒

Two Kinds of Methods of PCR for Detection of Influenza Virus

目的 建立特异敏感的快速检测流感病毒的分子生物学方法。方法 用多重逆转录聚合酶链反应和半巢式多重聚合酶链反应扩增甲1、甲3 和乙型流感病毒的HA基因的HA1片段。根据扩增产物的大小检测流感病毒。结果 以流感病毒的HA基因为模板设计的 3对引物能特异性地扩增出 94 2bp、1117bp和 75 1bp的核酸片段 ,它们分别和甲1,甲3 和乙型流感病毒有稳定对应关系。根据扩增产物的大小可以检测流感病毒。用这个方法能检测病毒浓度为0 1TCID50 的样品。结论 多重逆转录聚合酶链反应和半巢式多重聚合酶链反应是检测流感病毒的特异敏感的方法。

Objective To develop a specific and sensitive molecular biological method for rapid detection of influenza virus.Methods Multiplex reverse transcription-PCR and half nested multiplex-PCR were used to amplify and differentiate the hemagglutinin (HA) genes of influenza virus subtypes of H 1 and H 3 and type of B.Results Three primer pairs are specific for the influenza virus subtypes of H 1 and H 3 and type of B. And their HA gene amplicons are particular in size. The amplicon of H 1, H 3 and B are 942bp 1117bp and 751bp in size respectively.The type and subtypes of influenza virus can be detected according to the size of HA gene amplcon. The influenza virus can be detected in a 0.1 TCID \{50\} specimen.Conclusion Muitiplex reverse transcription-PCR and half nested multiplex-PCR are specific and sensitive molecular biological methods for rapid detection of influenza virus.