CAJ | 학술논문

目的建立施马伦贝格病毒逆转录环介导等温扩增(RT-LAMP)快速检测方法.方法从GenBank数据库中获得施马伦贝格病毒S基因序列,应用DNAStar软件MegAlign程序分析其序列,利用PrimerExplorerV4软件在序列保守区域设计LAMP引物,即外引物、内引物,建立施马伦贝格病毒等温扩增快速检测方法.结果 RT-LAMP检测方法对施马伦贝格病毒RNA的灵敏度达到10个拷贝,所用引物对赤羽病毒、口蹄疫病毒、蓝舌病毒、水泡性口炎病毒及牛病毒性腹泻病毒无特异性扩增,表现出良好特异性.结论建立的施马伦贝格病毒等温扩增快速检测方法灵敏度高、特异性强,为快速检测施马伦贝格病毒提供了新方法.
목적 건립시마륜패격병독역전록배개도등온확증(RT-LAMP)쾌속검측방법.방법 종GenBank수거고중획득시마륜패격병독S기인서렬,응용DNAStar연건MegAlign정서분석기서렬,이용PrimerExplorerV4연건재서렬보수구역설계LAMP인물,즉외인물、내인물,건립시마륜패격병독등온확증쾌속검측방법.결과 RT-LAMP검측방법대시마륜패격병독RNA적령민도체도10개고패,소용인물대적우병독、구제역병독、람설병독、수포성구염병독급우병독성복사병독무특이성확증,표현출량호특이성.결론 건립적시마륜패격병독등온확증쾌속검측방법령민도고、특이성강,위쾌속검측시마륜패격병독제공료신방법.
Objective To set up loop-mediated isothermal amplification(RT-LAMP) method for quick detection of Schmallenbergvirus.Methods The S gene sequences of Schmallenbergvirus were downloaded from GenBank database and analyzed using MegAlign tools of software DNAStar.The outer primers and inner primers of LAMP were then designed according to their conservative region using software PrimerExplorer V4,and the reaction conditions were optimized.Results The results showed that the RTLAMP assay had a detection limit of 10 RNA copies.The primers used in this assay showed the specificity for Schmallenberg virus and did not amplify FMDV,BVDV,BTV,AKAV and VSV.Conclusion It is indicated the RT-LAMP method is a simple,rapid,accurate,sensitive and specific method for detecting Schmallenberg virus.This assay offers a new method and idea for detecting Schmallenberg virus.