变异猪繁殖与呼吸综合征病毒RT-PCR检测方法的建立

Establishment of the Variant Porcine Reproductive and Respiratory Syndrome Virus by RT-PCR Assay

根据GenBank已发表及笔者所在研究室分离鉴定的变异猪繁殖与呼吸综合征病毒的NSP2基因序列,设计并合成了一对引物,建立了一种鉴别PRRSV的RT-PCR检测方法。结果表明:该方法扩增变异PRRSV基因组时可获得217bp的片段,扩增普通PRRSV时则获得307bp的片段,同条件扩增猪瘟病毒(CSFV)、圆环病毒Ⅱ型(PCV2)、伪狂犬病病毒(PRV)均为阴性;该体系可检测到2.6pg的PRRS病毒核酸,具有较高敏感性。对2007—2008年送检的40份临床样品进行检测,结果检出12份为变异PRRSV,其阳性率为30.0%。研究结果表明,建立的RT-PCR鉴别变异PRRSV方法具有特异性强、敏感性高、重复性好等特点,适用于对猪繁殖与呼吸综合征病毒的鉴别诊断。

A pair of primers was designed, based on the NSP2 gene sequence of PRRSV published in Genbank and RT-PCR assay was developed to distinguish common PRRSV and variant PRRSV which was isolated from Fujian province. 217 bp specifically fragment and 307bp fragment were amplified from variant PRRSV and classical form respectively. Negative results were found on CSFV, PCV2 and PRV. The developed method was sensitive to detect the RNA as low as 2.6pg. 40 clinical specimens of sick pigs were tested by the method from 2007 to 2008, 30 percent of them were positive of and variant PRRSV.variant PRRSV. The results showed that RT-PCR assay is high specificity, sensitivity and reproducibility, and the developed method could be used to differentiate between common PRRSV.