A method for simultaneous detection and identification of Brazilian dog- and vampire bat-related rabies virus by reverse transcription loop-mediated isothermal amplification assay
A method for simultaneous detection and identification of Brazilian dog- and vampire bat-related rabies virus by reverse transcription loop-mediated isothermal amplification assay
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DOI：
10.1016/j.jviromet.2010.04.008
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发表时间：
2010-09-01
影响因子：
3.1
通讯作者：
Sakai, Takeo
中科院分区：
文献类型：
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作者：
Saitou, Yasumasa;Kobayashi, Yuki;Sakai, Takeo
目前，在巴西发生的人类狂犬病散发病例可主要归因于与狗和吸血蝙蝠有关的狂犬病病毒。采用逆转录环介导的等温扩增（RT-LAMP）方法在60 min内同时检测两种狂犬病野变异体。利用限制性内切酶Alwl消化RT-LAMP反应的扩增子，可以将吸血蝙蝠相关的狂犬病病毒与狗的狂犬病病毒区分开来。扩增和酶切均可在提取RNA后120 min内完成。此外，RI-LAMP检测还在巴西食果蝙蝠和乌干达狗、牛和山羊样本分离物中检测到狂犬病毒。相比之下，一些巴西食虫蝙蝠和所有中国狗、猪和牛样本使用RI-LAMP检测结果为假阴性。本研究表明，RT-LAMP法可快速检测巴西狂犬病病毒原库分离株。为了使RT-LAMP检测能够用于野外狂犬病病毒遗传变异的普遍检测，还需要进一步改进。(C) 2010 Elsevier B.V.版权所有
At present, the sporadic occurrence of human rabies in Brazil can be attributed primarily to dog- and vampire bat-related rabies viruses. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) was employed as a simultaneous detection method for both rabies field variants within 60 min. Vampire bat-related rabies viruses could be distinguished from dog variants by digesting amplicons of the RT-LAMP reaction using the restriction enzyme Alwl. Amplification and digestion could both be completed within 120 min after RNA extraction. In addition, the RI-LAMP assay also detected rabies virus in isolates from Brazilian frugivorous bats and Ugandan dog, bovine and goat samples. In contrast, there were false negative results from several Brazilian insectivorous bats and all of Chinese dog, pig, and bovine samples using the RI-LAMP assay. This study showed that the RT-LAMP assay is effective for the rapid detection of rabies virus isolates from the primary reservoir in Brazil. Further improvements are necessary so that the RT-LAMP assay can be employed for the universal detection of genetic variants of rabies virus in the field. (C) 2010 Elsevier B.V. All rights reserved.