Rapid and simple detection of Ebola virus by reverse transcription-loop-mediated isothermal amplification
Rapid and simple detection of Ebola virus by reverse transcription-loop-mediated isothermal amplification
复制标题
DOI：
10.1016/j.jviromet.2006.11.031
复制
发表时间：
2007-04-01
影响因子：
3.1
通讯作者：
Yasuda, Jiro
中科院分区：
文献类型：
--
作者：
Kurosaki, Yohei;Takada, Ayato;Yasuda, Jiro
埃博拉病毒（EBOV）在人类和非人灵长类动物中引起严重的出血热，死亡率很高。需要快速识别病毒以防止感染扩散。在这项研究中，我们开发和评估了一个简单的一步逆转录环介导的等温扩增（RT-LAMP）检测扎伊尔埃博拉病毒（ZEBOV），最致命的EBOV种，针对病毒基因组的尾部区域的快速检测。该检测方法可以在26 min内检测到20个拷贝的人工ZEBOV RNA，具有真实的时间监测检测，并且还可以检测到10（-3）FFU的细胞培养繁殖的病毒。检测104 FFU的ZEBOV所需的反应时间仅为20 min。此外，该测定法对ZEBOV具有高度特异性。本研究建立的RT-LAMP检测方法具有快速、简便、特异性强、灵敏度高等优点，有望成为ZEBOV的有效诊断工具。此外，由于这种技术不需要复杂的仪器，它似乎非常适合在埃博拉疫情地区（如中非）的现场或实验室进行诊断。(c)2006 Elsevier B.V.保留所有权利。
Ebola virus (EBOV) causes severe hemorrhagic fever in humans and nonhuman primates with high mortality rates. Rapid identification of the virus is required to prevent spread of the infection. In this study, we developed and evaluated a one-step simple reverse transcription-loop mediated isothermal amplification (RT-LAMP) assay for the rapid detection of Zaire ebolavirus (ZEBOV), the most virulent species of EBOV, targeting the trailer region of the viral genome. The assay could detect 20 copies of the artificial ZEBOV RNA in 26 min with a real time-monitoring detection, and also detect 10(-3) FFU of the cell-culture propagated viruses. The reaction time needed to detect 104 FFU of ZEBOV was only 20 min. In addition, the assay was highly specific for ZEBOV. The RT-LAMP assay developed in this study is rapid, simple, highly specific, and sensitive for the detection of ZEBOV, and so may be an effective diagnostic tool. Furthermore, as this technique does not require sophisticated instrumentation, it seems very suitable for diagnosis in the field or laboratories in Ebola outbreak areas such as Central Africa. (c) 2006 Elsevier B.V. All rights reserved.