Development of a loop-mediated isothermal amplification method for rapid detection of porcine boca-like virus

文献类型： 外文期刊

第一作者： Li, Bin

作者： Li, Bin;Ma, Jun-jie;Zhang, Xue-han;Wen, Li-bin;Mao, Li;Ni, Yan-xiu;Guo, Rong-li;Zhou, Jun-ming;Lv, Li-xin;He, Kong-wang;Li, Bin;Ma, Jun-jie;Zhang, Xue-han;Wen, Li-bin;Mao, Li;Ni, Yan-xiu;Guo, Rong-li;Zhou, Jun-ming;Lv, Li-xin;He, Kong-wang;Li, Bin;Ma, Jun-jie;Zhang, Xue-han;Wen, Li-bin;Mao, Li;Ni, Yan-xiu;Guo, Rong-li;Zhou, Jun-ming;Lv, Li-xin;He, Kong-wang;Xiao, Shao-bo

作者机构：

关键词： Porcine boca-like virus;Loop-mediated isothermal amplification (LAMP);PCR;Detection

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： 2012 年 179 卷 2 期

页码：

收录情况： SCI

摘要： The porcine boca-like virus (Pbo-likeV) was recently discovered in Swedish pigs with post-weaning multisystemic wasting syndrome (PMWS). In this study, a loop-mediated isothermal amplification (LAMP) assay was developed for rapid, specific and sensitive detection of Pbo-likeV. A set of four primers specific for six regions of Pbo-likeVVP1/2 genes was designed with the online software. The reaction temperature and time were optimized to 65 degrees C and 60 min, respectively. LAMP products were detected by agarose gel electrophoresis or by visual inspection of a color change due to addition of fluorescent dye. The developed method was highly specific for detection of Pbo-likev, and no cross-reaction was observed with other swine viruses, such as porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), porcine parvovirus (PPV) and classic swine fever virus (CSFV) found commonly in China. The lower detection limit of the LAMP assay was approximately 10 copies per reaction, and it was 100 times more sensitive than that of conventional PCR. Furthermore, the efficiency of LAMP for detection Pbo-likeV in clinical samples was comparable to PCR and sequencing. These results showed that the LAMP assay is a simple, rapid, sensitive and specific technique for detection of Pbo-likeV, and the procedure of LAMP does not rely on any special equipment. It has capacity for the detection of Pbo-likeV both in the laboratory and on farms. (C) 2011 Elsevier B.V. All rights reserved.

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