摘要
根据猪瘟病毒(CSFV)P120基因及高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)Nsp2基因缺失区两端的保守区分别设计合成1对特异性引物,建立了一种简便、快速、同时鉴别检测CSFV、PRRSV、HP-PRRSV的多重RT-PCR方法。该方法扩增CSFV基因组时可获得508bp的片段,扩增PRRSV时可获得320bp的片段,扩增HP-PRRSV基因组时可获得230bp的片段,因此,根据多重RT-PCR产物大小可将三者区分开来。利用建立的多重RT-PCR方法,对50份临床可疑病料进行检测,结果发现,CSFV阳性率为8%,PRRSV阳性率为18%,HP-PRRSV阳性率为22%,而CSFV和HP-PRRSV混合感染阳性率达2%,PRRSV和HP-PRRSV混合感染阳性率达16%,说明所建立的多重RT-PCR方法简便、快速、特异,可同时鉴别CSFV、PRRSV及HP-PRRSV,亦可用于感染这3种病毒的临床病料的快速鉴定和分子流行病学调查。
The present study was conducted to establish a simple and rapid multiplex RT-PCR method for simultaneous detection of classical swine fever virus（CSFV） and classical and highly pathogenic porcine reproductive and respiratory syndrome（PRRSV）.A pairs of primers was synthesized,respectively,according to P120 gene for CSFV and the gene of two non-continued deletions in the Nsp2 region of viral genome for HP-PRRSV.A 508 bp fragment was obtained specifically from amplification of CSFVs,a 230 bp fragment was obtained from the highly pathogenic PRRSVs and a 320 bp fragment was obtained from the classical form of PRRSVs.Different RT-PCR amplification products differentiated three pathogens,efficiently.50 samples collected from the diseased pigs and aborted fetuses were detected by multiplex RT-PCR method,out of which 8% were found positive for CSFV,and 18% and 22% for classical and HP-PRRSV,respectively.Further,2% and 16% positive co-infection results for CSFV / HP-PRRSV and HP-PRRSV / the classical PRRSV had also been detected.These results indicated the simplicity,rapidness and specificity of multiplex RT-PCR method in detection of these viruses.It could be used as an effective tool for differentiation,diagnosis and epidemiological investigations of clinical sam-ples infected by CSFV,HP-PRRSV and the classical PRRSV.
出处
《广西农业科学》
CSCD
2009年第11期1476-1480,共5页
Guangxi Agricultural Sciences
基金
广西柳州市科技攻关项目(2008060605)
