多重逆转录聚合酶链反应检测流感病毒

MULTIPLEX REVERSE TRANSCRIPTION-PCR FOR DETECTING 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连接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基因为模板设计的三对引物能特异性地扩增出 94 2 bp、 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 .1TCID5 0的样品。还可直接从疑似流感病人的含漱液中检测到乙型流感病毒。结论 :多重逆转录聚合酶链反应连接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,rapid and sensitive molecular biological method for detecting influenza virus. Method:Multiplex reverse transcription-PCR and multiplex-PCR were used to amplify and differentiate the hemagglutinin (HA) genes of H 1,H 3 and B influenza virus.Result:Three primer pairs were specific to H 1?H 3 and B influenza virus. And their HA gene amplicons were particular in size. The amplicon of H 1?H 3 and B were 942bp?1117bp and 751bp in size,respectively. Influenza virus could be detected according to the size of HA gene amplicon. The type of influenza virus B has been detected in clinical specimen by multiplex reverse transcription-PCR and multiplex-PCR. Conclusion:Multiplex reverse transcription-PCR and multiplex-PCR were specific,rapid and sensitive molecular biological methods for detecting influenza virus. The type of influenza virus B could be detected in clinical specimen by multiplex reverse transcription-PCR and multiplex-PCR.