Visual detection of pandemic influenza A H1N1 Virus 2009 by reverse-transcription loop-mediated isothermal amplification with hydroxynaphthol blue dye.

Xue-Jun Ma, Yue-Long Shu, Kai Nie, Meng Qin, Da-Yan Wang, Rong-Bao Gao, Miao Wang, Le-Ying Wen, Feng Han, Shu-Mei Zhou, Xiang Zhao, Yan-Hui Cheng, De-Xin Li, Xiao-Ping Dong

Index: J. Virol. Methods 167(2) , 214-7, (2010)

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Abstract

A sensitive reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of pandemic influenza A H1N1 virus infection. The reaction was performed in one step in a single tube at 65 degrees C for 60 min with the addition of hydroxynaphthol blue (HNB) dye prior to amplification. The detection limit of the RT-LAMP assay was approximately 60 copies, and no cross-detection was observed. The assay was evaluated further with 50 clinical specimens diagnosed clinically with seasonal influenza or pandemic influenza A H1N1 virus infection. RT-LAMP with HNB dye was demonstrated to be a sensitive and easy assay for rapid detection of pandemic influenza A H1N1 virus.Copyright (c) 2010 Elsevier B.V. All rights reserved.