猪细环病毒1型环介导等温扩增方法的建立

Establishment of a Loop-mediated Isothermal Amplification Assay for Detection of Torque Teno Sus Virus 1

猪细环病毒(Torque teno sus virus,TTSu V)是一种无囊膜,单链的DNA病毒,存在猪细环病毒1型和猪细环病毒2型2个亚型。为建立一种新型快速的TTSu V1检测方法。针对TTSu V1保守的非编码区域(untranslated regions,UTR)设计2对LAMP引物,并对反应体系和反应条件进行了优化调整,建立了TTSu V1 LAMP检测方法。结果表明该方法最佳反应条件为62℃1 h,病毒最低检出限可达9.2 copies/μL,特异性良好。LAMP反应是一种集特异性强,灵敏度高,反应速度快于一体的的基因检测方法,有望成为快速检测TTSu V1的手段之一。

Torque teno sus virus （TFSuV） is a non-enveloped, single-stranded DNA virus which has two subtypes.The loop-mediated isothermal amplification （LAMP） for detecting TrSuV1 was established with a set of primers designed according to the conserved untranslated regions （UTR） of TFSuV1.The results showed that the optimized reaction conditions of LAMP for detecting TTSuV1 were in a constant temperature at 62 ℃ for 60 min.The method was able to detect TTSuV1 with specificity and could achieve a detection limit of 9.2 copies/ μL TTSuV1 DNA molecules.The LAMP method established in this study is a rapid, sensitive, specific assay and it is expected to become a tool for detection of TTSuV1.