| 用反转录-聚合酶链反应检测猪繁殖与呼吸综合征病毒的研究 |
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| 引用本文： | 周婷,文心田,曹三杰,肖驰,王新,张雪锋.用反转录-聚合酶链反应检测猪繁殖与呼吸综合征病毒的研究[J].四川农业大学学报,2005,23(2):244-246,252. |
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| 作者姓名： | 周婷 文心田 曹三杰 肖驰 王新 张雪锋 |
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| 作者单位： | 四川农业大学,动物科技学院,四川,雅安,625014 |
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| 摘 要： | 根据猪繁殖与呼吸综合征病毒(PRRSV)美洲型毒株的基因序列设计了一对特异性引物P1/P2,扩增的片段为550bp,根据欧洲型毒株设计了一对特异性检测引物P3/P4,扩增的片段为433bp,建立PRRSV的RT-PCR的检测与血清型鉴别方法。特异性试验表明,这2对引物均不能扩增其他常见的繁殖障碍相关病毒的RNA或DNA。敏感性试验表明,这2对引物可以分别到检测10-5TCID50和10-4TCID50的病毒含量。利用该方法对重庆、四川某些猪场中临床上疑似为PRRS的20份送检组织样品进行检测,只有引物P1/P2能扩增出与预期大小相符的RT-PCR产物;其中有15份样品呈PRRSV阳性结果,阳性率为75%,说明送检的样品感染的PRRSV为美洲型。试验结果表明该方法快速检测病料组织中PRRSV是可行的,是临床上对PRRS进行快速诊断和分子流行病学调查的实用方法。
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| 关 键 词： | 猪繁殖与呼吸综合征病毒 反转录-聚合酶链反应 检测 |
| 文章编号： | 1000-2650(2005)02-0244-03 |
Studies on the Detection of Porcine Reproductive and Respiratory Syndrome Virus by RT-PCR |
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| ZHOU Ting,WEN Xin-tian,CAO San-jie,XIAO Chi,WANG Xin,ZHANG Xue-feng.Studies on the Detection of Porcine Reproductive and Respiratory Syndrome Virus by RT-PCR[J].Journal of Sichuan Agricultural University,2005,23(2):244-246,252. |
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| Authors: | ZHOU Ting WEN Xin-tian CAO San-jie XIAO Chi WANG Xin ZHANG Xue-feng |
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| Institution: | ZHOU Ting,WEN Xin-tian~,CAO San-jie,XIAO Chi,WANG Xin,ZHANG Xue-feng |
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| Abstract: | According to the GenBank published sequence of porcine reproductive and respiratory sydrome virus (PRRSV), P1/P2 primes were designed to amplify the American-type gene (about 550?bp), and P3/P4 primes were designed to amplify the European-type gene (about 430?bp). In the research, we established the identifying and distinguishing PRRSV RT-PCR method with the P1/P2 and P3/P4 primes. There was no amplification observed when using RNA or DNA extracted from pseudorabies virus (PRV), Porcine parvovirus (PPV) and Classical Swine fever virus (CSF), which showed the assay had good specificity. And the assay was also highly sensitive and could detected as little as 10~ -5 TCID_ 50 and 10~ -4 TCID_ 50 of virus template. The lung, spleen and lymphoid tissues of 20 doubted-to-be-infected-with-PRRSV pigs from the pig farms in Chongqing and Sichuan were detected by RT-PCR. A gene fragment about 550?bp was amplified from 15 of 20 samples, while a gene fragment about 430?bp was not amplified, the results confirmed the amplified mixed tissues belonged to Americana type. The RT-PCR was proved to be a specific, sensitive, rapid and simple method and could be used to research on epidemiology and early clinical diagnosis of PRRS in molecular level. |
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| Keywords: | PRRSV RT-PCR detection |
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