Direct and rapid detection of porcine epidemic diarrhea virus by RT-PCR
Direct and rapid detection of porcine epidemic diarrhea virus by RT-PCR
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DOI：
10.1016/s0166-0934(97)00157-2
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发表时间：
1997-12-01
影响因子：
3.1
通讯作者：
Suzuki, S
中科院分区：
文献类型：
--
作者：
Ishikawa, K;Sekiguchi, H;Suzuki, S
为了建立一种实用的猪流行性腹泻病毒（PEDV）检测方法，研究了利用PEDV M蛋白基因序列设计引物，建立PEDV的RT-PCR检测体系。设计引物，通过RT-PCR扩增出854 bp的目的片段。该反应对PEDV RNA是特异性的，但对所测试的其它病毒属不是特异性的。在用PEDV与小肠或粪便匀浆的混合物进行的实验中，该方法可以在8小时内有效地从含有非常低数量的病毒（100 TCID 50/样品）的样品中检测PEDV RNA。从发病猪场采集的12份标本中，有4份肠道标本检出PEDV RNA。结果表明，RT-PCR检测PEDV及其抗原的结果与病毒分离和链霉亲和素-生物素技术检测PEDV及其抗原的结果一致，表明RT-PCR检测PEDV及其抗原具有实用价值。(C)1997年Elsevier Science B.V.
To establish a practical method for detecting porcine epidemic diarrhea virus (PEDV), the use of primers derived from sequences that amplify the M protein genes of PEDV in a RT-PCR detection system was investigated. Primers were designed to amplify a 854-bp fragment by RT-PCR. This reaction was specific to the PEDV RNA but not to that of other viral genera tested. In experiments with mixtures of PEDV and either small intestine or fecal homogenates, this method could detect efficiently the PEDV RNA from samples containing very low numbers of virus (100 TCID50/sample) within 8 h. With specimens collected from swine breeding farms with the diarrhoeal disease, the PEDV RNA was detected in four intestine specimens out of II specimens. The result was in close agreement with the results of virus isolation and streptavidin-biotin technique for detecting PEDV and its antigens, suggesting that the RT-PCR assay would be useful method for practical application. (C) 1997 Elsevier Science B.V.