摘要
目的建立特异敏感的快速检测含漱液中的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ＨＡ基因的ＨＡ１片段，得到各病毒与特异性大小的扩增产物的对应关系。用同样的方法扩增含漱液中流感病毒的ＨＡ基因的ＨＡ１片段。根据这对应关系检测含漱液中的流感病毒。结果以流感病毒的ＨＡ基因为模板设计的三对引物，用多重逆转录聚合酶链反应能特异性地扩增出９４２ｂｐ、１１１７ｂｐ和 ７５１ｂｐ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。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含漱液中流感病毒ＨＡ基因的ＨＡ１片段，根据扩增产物的大小检出含漱液中的乙型流感病毒。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能特异敏感地检出含漱液中的乙型流感病毒。
objective To develop a specific and sensitive molecualar biological method for.rapid detection of influenza virus in clinical specimens Method multiplex reverse transcrption-PCR and half nested multiplex PCR were used to amplify and differentiate the hem- agglutinin (HA)genes of influenza virus subtyps of H1 and H3and type of B.The HA gene of influenza in clinical specimens was aplied by multiplex reverse transcription-PCR.The influenza virus in clinical specimen could bedetected according to the size of amplicons.Re- sult Three primer pairs are specific for the influenza virus subtypes of H1 and type of B.And their HA gene amplicons are par- ticular in size.The amplicons of H1,H3 and B are 942bp, 1117bp and 751bp in size respectively.The type and subtypes of influenza virus can be detected in clinical specimen according to the size of HA gene amplicon.Conclusion The type of influenza virus B can be rapid detected specifically and sensitively in clinical specimen by multiplex reverse transcription-PCR and
出处
《中国卫生检验杂志》
CAS
2001年第1期6-8,共3页
Chinese Journal of Health Laboratory Technology
