Imai Masaki, Ninomiya Ai, Minekawa Harumi, Notomi Tsugunori, Ishizaki Toru, Tashiro Masato, Odagiri Takato
Laboratory of Influenza Viruses, Department of Virology 3, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-Murayama, Tokyo 208-0011, Japan.
Vaccine. 2006 Nov 10;24(44-46):6679-82. doi: 10.1016/j.vaccine.2006.05.046. Epub 2006 Jun 8.
We developed a rapid and sensitive diagnosis system for H5N1 highly pathogenic avian influenza (HPAI) virus infection using an unique gene amplification method, reverse transcriptase loop-mediated isothermal amplification (RT-LAMP). The sensitivity of the system was found to be 100-fold higher than that of ordinary one-step RT-PCR. Moreover, by using viral RNAs extracted from influenza viruses of all 15 HA subtypes, the RT-LAMP system was confirmed to amplify only the RNA of H5 subtype virus. In the surveillance of H5N1 virus infection of wild birds, we detected two positive cases from dead crows found near the affected area with H5N1-HPAI by using RT-LAMP system, although one of two positive cases was missed by RT-PCR. These results suggested that our newly developed RT-LAMP system specific for H5 virus would be a beneficial diagnostic tool for surveillance of recent outbreaks caused by H5N1-HPAI viruses.
我们利用一种独特的基因扩增方法——逆转录环介导等温扩增（RT-LAMP），开发了一种用于H5N1高致病性禽流感（HPAI）病毒感染的快速灵敏诊断系统。该系统的灵敏度比普通一步法RT-PCR高100倍。此外，通过使用从所有15种HA亚型流感病毒中提取的病毒RNA，证实RT-LAMP系统仅能扩增H5亚型病毒的RNA。在对野生鸟类H5N1病毒感染的监测中，我们使用RT-LAMP系统从受H5N1-HPAI影响区域附近发现的死乌鸦中检测到两例阳性病例，尽管RT-PCR漏检了两例阳性病例中的一例。这些结果表明，我们新开发的针对H5病毒的RT-LAMP系统将是监测由H5N1-HPAI病毒引起的近期疫情的有益诊断工具。
