福建省猪群感染牛病毒性腹泻病毒的病原学检测与分析

Investigation of Bovine Viral Diarrhea Virus(BVDV)from Swine in Fujian Province

旨在建立鉴别牛病毒性腹泻病毒(BVDV)与猪瘟病毒(CSFV)的RT-PCR检测方法,用于福建省猪群感染BVDV的病原学检测,并进行猪源BVDV协同感染的病原学调查。根据BVDV 5′-UTR保守序列建立猪源BVDV的特异性RT-PCR,进行敏感性、特异性、重复性试验,然后对福建省9个地市42个猪场的458份临床样品进行BVDV检测及测序,并参照已有的CSFV、PRRSV、HP-PRRSV、PRV和PCV-2病原检测方法对BVDV阳性样品进行协同感染检测。结果显示该方法敏感度达0.1 TCID50,检测CSFV、PRRSV、HP-PRRSV、PRV、PCV-2和PPV均为阴性,重复5次进行敏感性和特异性试验的结果一致;样品BVDV病原阳性率10.04%(46/458),猪场阳性率78.57%(33/42),9个地市阳性率100%(9/9);其中46份BVDV阳性样品CSFV、PRRSV、HP-PRRSV、PRV和PCV-2感染的阳性率分别为91.30%(42/46)、50.00%(23/46)、41.30%(19/46)、63.04%(29/46)、37.00%(17/46)。结果表明,建立的RT-PCR方法敏感性高、特异性强、重复性好,适合用于猪源BVDV的监测;福建省猪群BVDV感染情况较普遍,且与多种病毒协同,其中CSFV协同感染率最高。

The study was aimed at establishing a reverse transcriptase PCR(RT-PCR)to differentiate swine bovine viral diarrhea virus(BVDV)and classical swine fever virus(CSFV)for the investigation of BVDV from swine in Fujian province and combined infection of swine BVDV.According to the conserved gene sequence of BVDV 5′-UTR,a specific RT-PCR was established and then sensitivity,specificity,repeatability were tested for the detection of BVDV in 458 clinical samples from 42 swine farms in 9regions of Fujian province.Then according to the reference CSFV,porcine reproductive and respiratory syndrome virus(PRRSV),highly pathogenic PRRSV(HP-PRRSV),pseudorabies virus(PRV)and porcine circovirus type 2(PCV-2)were detected in BVDV positive samples.The results showed that sensitivity was determined as 0.1 TCID50 BVDV.In addition,all negative controls such as CSFV,PRRSV,HP-PRRSV,PRV,PCV-2and porcine parvovirus virus(PPV)showed negative detection in the specificity assay.Both sensitivity and specificity were repeated five times with similar results.The BVDV positive rate of swine,swine farm and 9regions was 10.04%(46/458),78.57%(33/42)and 100%(9/9)in samples by sequence data,respectively,and the positive rate of 46 BVDV positive samples which infected with CSFV,PRRSV,HP-PRRSV,PRV and PCV-2were 91.30%(42/46),50.00%(23/46),41.30%(19/46),63.04%(29/46)and 37.00%(17/46).These findings indicated that the method with high sensitivity,specificity and reproducibility could provide an efficient tool for detection of swine BVDV.The BVDV infection in swine was common in Fujian province.And it was combined infection with other pathogens,especially CSFV.