Rapid detection of duck hepatitis A virus genotype C using reverse transcription loop-mediated isothermal amplification

文献类型： 外文期刊

第一作者： Li, Chuanfeng

作者： Li, Chuanfeng;Chen, Zongyan;Meng, Chunchun;Liu, Guangqing

作者机构：

关键词： Duck hepatitis A virus;Loop-mediated isothermal amplification;RNA-dependent RNA polymerase

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： 2014 年 196 卷

页码：

收录情况： SCI

摘要： A one-step reverse transcription loop-mediated isothermal amplification (RI-LAMP) assay was used and optimized to develop a rapid and sensitive detection system for duck hepatitis A virus genotype C (DHAV-C) RNA. A set of four specific primers was designed against highly conserved sequences located within the 3D gene from DHAV (strain GX1201). Under optimal reaction conditions, the sensitivity of DHAV-C-specific RI-LAMP was 100-fold higher than that of reverse transcriptase-polymerase chain reaction (RT-PCR), with a detection limit of 0.3 pg (6.59 x 104 copies) per reaction. No cross-reactivity was observed from the samples of other duck viruses, which is in good accordance with RT-PCR. Furthermore, a positive reaction can be visually inspected by observing turbidity or color change after the addition of SYBR green I dye. The DHAV-C-specific RT-LAMP assay was applied to the samples and compared with RT-PCR. The positive-sample ratios were 26.7% (12 of 45) by RT-LAMP and 20% (9 of 45) by RT-PCR. Therefore, the newly developed RT-LAMP assay is a rapid, specific, sensitive, and cost-effective method of DHAV-C detection. This assay has potential applications in both clinical diagnosis and field surveillance of DHAV-C infection. (C) 2013 Published by Elsevier B.V.

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