家蝇体内猪繁殖与呼吸综合征病毒的初步分离与鉴定

为调查养猪场环境中家蝇携带猪繁殖与呼吸综合征病毒(PRRSV)情况,2010年采集贵州某猪场家蝇样本,采用RT-PCR方法筛选阳性样本,接种Marc-145细胞分离培养病毒,对分离获得的家蝇样本的PRRSV Nsp2基因进行克隆和序列分析。针对PRRSV N基因家蝇样本扩增出377 bp的特异性片段,阳性家蝇样本接种的Marc-145细胞出现明显CPE现象,针对PRRSV Nsp2基因细胞培养物扩增出1064 bp的特异性片段,测序结果显示,家蝇样本的细胞培养物的PRRSV Nsp2基因片段与贵州猪场猪源PRRSV流行株相比具有较高的核苷酸同源性,高达97.1%~98.5%。从而初步推断家蝇也可能成为PRRSV携带者。

Isolation and Identification of Porcine Reproductive and Respiratory Syndrome Virus in the Indigenous Flies

To survey the indigenous flies to carry porcine reproductive and respiratory syndrome virus(PRRSV) in the porcine farm,in 2010,the indigenous flies in the Guizhou porcine farm were collected to detect the pathogeny of PRRSV by RT-PCR.The positive indigenous flies were chosen to isolate and cultivate the virus by inoculating in the cells of Marc-145,and the genes of the isolated PRRSV were cloned and analysed by testing the sequences.The findings indicate that the length of the PRRSV N gene fragments in the flies is 377 bp and the cytopathic effects appear in the cells of Marc-145.The cells were inoculated by the PRRSV.The length of the PRRSV Nsp2 gene fragments in the cells is 1064 bp.By analyzing the consequences of the sequences of the PCR products that were cloned,the findings indicate that the fragment of PRRSV Nsp2 gene in the cells that inoculated by the virus of PRRS and in the swine of Guizhou province are similar and the rate of the similarity is 97.1%—98.5%.So the initial inference is that the indigenous flies may be the carriers of PRRSV.