Development of a reverse-transcription loop-mediated isothermal amplification method for detection of rabbit hemorrhagic disease virus

文献类型： 外文期刊

第一作者： Yuan, Dongwei

作者： Yuan, Dongwei;Guo, Dongchun;Liu, Jiasen;Si, Changde;Jiang, Qian;Lin, Huan;Yang, Tiankuo;Qu, Liandong;Yuan, Dongwei

作者机构：

关键词： Rabbit hemorrhagic disease virus;Reverse transcription;Loop-mediated isothermal amplification;Sensitivity;Reliability

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN： 0166-0934

年卷期： 2013 年 187 卷 2 期

页码：

收录情况： SCI

摘要： Rabbit hemorrhagic disease virus (RHDV) causes haemagglutination and severe liver damage, with a high mortality rate. To develop a rapid and sensitive method for the surveillance of RHDV, a one-step reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay was established using a set of four primers specific for the VP60 gene segment of RHDV. The established assay was performed at 64 degrees C for 40 min under isothermal conditions, and the results were visualized directly by electrophoresis or as fluorescent signals under ultraviolet light. The detection limit of the RI-LAMP assay was 10 copies of viral RNA per reaction, which was comparable to quantitative real-time RT-PCR, and 100-fold more sensitive than standard RT-PCR. Furthermore, seven viral RNAs of field isolates in China could be detected successfully using this assay. Overall, the newly established RT-LAMP assay indicates the potential usefulness of the technique as a simple, rapid and sensitive procedure, and can visually detect RHDV infection without the need for any specialized equipment. (C) 2012 Elsevier B.V. All rights reserved.

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