摘要
参照GenBank的变异型猪繁殖与呼吸综合征病毒(PRRSV)与普通PRRSV的Nsp2段基因序列,设计并合成1对引物和TaqMan探针。通过对反应条件优化,标准质粒构建,建立了TaqMan荧光定量RT-PCR诊断变异型PRRSV方法。结果表明,该方法具有特异性强,敏感性高等特点,能够检测出264个拷贝数的标准质粒品,0.562 3TICD50的病毒量,比RT-PCR敏感10倍。对22份病料样品进行检测,结果有8份为阳性,阳性率为36.4%。由于该方法具有定量、快速、准确、敏感等优点,适用于对猪群感染变异型猪繁殖与呼吸综合征(PRRS)早、中、晚期的诊断,对有效诊断及防治高致病性PRRS的发生起到重要作用。
A pair of primers and a TaqMan fluorogenic probe were designed by comparing with the Nsp2 gene sequences of the variability porcine reproductive and respiratory syndrome virus（PRRSV） and the normal PRRSV.A real-time RT-PCR assay for the detection of the variability PRRSV was developed by conditional optimize and construction of standard plasmid.The results showed that the real-time RT-PCR assay was specific,and no cross reaction to normal PRRSV and other common porcine viruses.The sensitivity was 0.562 5 TCID50 of the viruses and 264 copy/mL standard plasmid,which was 10 times higher than conventional RT-PCR assay.22 sceptical specimens detected by the real-time RT-PCR showed 36.4%（8 out of 22） positive.The method would be useful for the diagnosis and control of the highly virulent PRRSV.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2010年第8期1023-1027,共5页
Chinese Journal of Veterinary Science
基金
福建省自然科学基金资助项目(2007J0060)
福建省科技重点资助项目(2006N0083)
