PRRSV Nsp9蛋白基因的RT-LAMP检测方法的建立

Establishment of rapid method for detection PRRSV on Nsp9 by a reverse loopmediated isothermal amplification(RT-LAMP)

根据GenBank中登录的PRRSV Nsp9基因序列,应用Primer Explorer V4软件,在该基因序列中选取保守区设计了4条RT-LAMP引物,旨在建立1种针对Nsp9蛋白基因的逆转录环介导等温扩增(RT-LAMP)的快速检测方法。采用引物分组扩增的方法对引物进行了检测,以确保引物的可靠性。对反应体系、温度以及时间进行了优化,检测了该方法的特异性和灵敏度。结果表明,该方法在等温条件下只需50min就能检测出结果。与RT-PCR方法相比,在判定检测结果时不需要借助昂贵仪器设备,具有高特异性、高灵敏度、操作简便快速等特点,适合于临床检测的应用。

In this study, according to the published PRRSV Nsp9 sequences in GenBank we de- signed four primers by using PrimerExplorerV4 software. Aiming at establishing a rapid detec- tion of porcine reproductive and respiratory syndrome virus （PRRSV） by rcverse transcription- loop-mediated isothermal amplification assay （RT-LAMP）. First,the group detection method was used to ensure reliability of the primers. Then, reaction system and reaction time were optimized, specificity and sensitivity were detected. The result indicated that PRRSV could be detected with- in 50 minutes by this method,with ideal specificity, shorter reaction time and higher sensitivity than general RT-PCR. The method was easy to operation, which was suitable for rapid detection of PRRSV in clinic.