猪蓝耳病病毒通用RT-PCR检测方法的建立及应用

Establishment and Application of Universal RT-PCR Detective method for PRRS virus

关键词:  猪繁殖与呼吸综合征病毒  ORF6~7基因  RT-PCR

Key words:  Porcine reproductive and respiratory syndrome virus  ORF6~7 gene  RT-PCR

摘要:

参考Genbank发表的美洲型猪蓝耳病病毒（Porcine Reproductive and Respiratory Syndrone virus, PRRSV ) VR-2332、CH-1a、JXA1等毒株的的主要结构蛋白基因序列，设计了一对引物，扩增目的片段为690bp。进行了RT-PCR检测方法的特异性、敏感性、重复性试验，建立了美洲型PRRSV通用RT-PCR检测方法。应用此方法对2009年1月-12月收集的126份临床样品进行了检测，检出率达44.4﹪。符合性检测结果表明，该法比高致病性PRRSV检测方法具有更广的监测范围。结果表明该方法具有特异、敏感、重复性好等优点，可用于PRRSV的临床发病检测及流行病学监测等工作。

Abstract:

According to the ORF5~7 genomic sequence of porcine reproductive and respiratory syndrome virus which published in Genbank, and using the Primer5.0 software, a pair of primers were designed, the motive fragment of amplification is 690bp. An universal RT-PCR detective method was established by specificity, sensibility, reproducibility,The method had some advantages such as fast, specific, sensitive and repeatable. We have Used the method to detect 126 clinic samples that collected from January to December,2009, the positive rate was 44.4%. Conformance test results indicate that this method has a wider detected rang than the highly pathogenic PRRSV RT-PCR method.The results show that this method is specific, sensitive, repeatable and can be used to detect and epidemiological surveillance of American-type PRRSV, etc.