Hu Dan, Hao Lina, Zhang Jinhai, Yao Pingping, Zhang Qi, Lv Heng, Gong Xiufang, Pan Xiuzhen, Cao Min, Zhu Jin, Zhang Yun, Feng Youjun, Wang Changjun
Research Institute for Medicine of Nanjing Command, Nanjing 210002, China.
Zhejiang Center for Disease Control and Prevention, Hangzhou 310051, China.
J Virol Methods. 2015 Sep 1;221:68-73. doi: 10.1016/j.jviromet.2015.04.017. Epub 2015 Apr 25.
We developed two assays based on one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) to identify Hantaan virus (HTNV) and Seoul virus (SEOV), members of the Hantavirus genus that cause hemorrhagic fever with renal syndrome (HFRS). Our results showed that these assays can be conducted within 30min under isothermal conditions. The detection limit for HTNV was around 10 copies per reaction, similar to detection levels for quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays. The detection limit for SEOV was 100 copies per reaction, a sensitivity that was 10-fold lower than that for qRT-PCR assays but 10-fold higher than that for RT-PCR assays. The method we developed was specific for both HTNV and SEOV without any cross-reaction with other pathogens. We conclude that RT-LAMP assays could be useful for the rapid and direct detection of HTNV and SEOV clinically, and for the epidemiological investigation of HFRS.
我们基于一步法逆转录环介导等温扩增（RT-LAMP）开发了两种检测方法，以鉴定汉坦病毒属的汉滩病毒（HTNV）和汉城病毒（SEOV），这两种病毒可引起肾综合征出血热（HFRS）。我们的结果表明，这些检测方法可在等温条件下30分钟内完成。HTNV的检测限约为每个反应10个拷贝，与定量逆转录聚合酶链反应（qRT-PCR）检测水平相似。SEOV的检测限为每个反应100个拷贝，其灵敏度比qRT-PCR检测低10倍，但比RT-PCR检测高10倍。我们开发的方法对HTNV和SEOV均具有特异性，与其他病原体无任何交叉反应。我们得出结论，RT-LAMP检测方法可用于临床快速直接检测HTNV和SEOV，以及HFRS的流行病学调查。
