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新型鸭呼肠孤病毒RT-LAMP检测方法的建立

DOI: 10.13560/j.cnki.biotech.bull.1985.2015.08.011, PP. 71-75

Keywords: 新型鸭呼肠孤病毒,S3基因,一步反转录环介导等温扩增

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Abstract:

建立适于基层实验室快速检测新型鸭呼肠孤病毒(Novelduckreovirus,NDRV)的一步反转录环介导等温扩增(RT-LAMP)方法。基于新型鸭呼肠孤病毒S3基因的6个保守区域设计了4条LAMP引物,利用BstDNA聚合酶在63℃恒温保持45min即可完成反转录和扩增反应,由此建立了RT-LAMP检测方法。该方法具有良好的特异性,除NDRV外对其他6种常见鸭病的检测结果均为阴性。该方法对病毒RNA的最低检出量为0.1pg,是常规RT-PCR方法的100倍。临床应用结果表明,该方法与病毒分离鉴定方法的符合率为98%,而且对仪器的要求低,适于基层实验室和现场检测。

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