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Development of a loop-mediated isothermal amplification assay for the detection of porcine hokovirus

文献类型： 外文期刊

作者： Li, Bin 1 ; Sun, Bing 1 ; Du, Lu-ping 1 ; Mao, Ai-hua 1 ; Wen, Li-bin 1 ; Ni, Yan-xiu 1 ; Zhang, Xue-han 2 ; He, Kong-wan 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Key Lab Vet Biol Engn & Technol, Natl Ctr Engn Res Vet Bioprod, Inst Vet Med,Minist Agr, Nanjing 210014, Jiangsu, Peoples R China

2.Jiangsu Acad Agr Sci, Key Lab Vet Biol Engn & Technol, Natl Ctr Engn Res Vet Bioprod, Inst Vet Med,Minist

关键词： Porcine hokovirus;Loop-mediated isothermal amplification;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： 2013 年 193 卷 2 期

页码：

收录情况： SCI

摘要： Hokoviruses have recently been detected as pathogens belonging to the family Parvoviridae, which comprises porcine hokovirus (PHoV) and bovine hokovirus (BHoV). In this study, we developed a loopmediated isothermal amplification (LAMP) assay for the rapid, specific and sensitive detection of PHoV. A set of four primers specific for six regions within the PHoV VP1/2 genes was designed using online software. The reaction temperature and time were optimized at 65 degrees C and 60 min, respectively. LAMP products were detected by agarose gel electrophoresis or by visual inspection of a color change caused by a fluorescent dye. The method was highly specific for PHoV, and no cross-reaction was observed with porcine circovirus type 2 (PCV2), porcine parvovirus (PPV), porcine bocavirus (PBoV), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV), classic swine fever virus (CSFV), or Japanese encephalitis virus (JEV). The detection limit was approximately 10 copies per reaction, which was 10 times more sensitive than conventional PCR. Furthermore, the efficiency of detection of PHoV in clinical samples was comparable to that of PCR and sequencing. These results show that the LAMP assay is a simple, rapid, sensitive and specific method for detecting PHoV. It does not require specialized equipment and can be used to detect PHoV both in the laboratory and in the field. (C) 2013 Elsevier B.V. All rights reserved.

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