猪繁殖与呼吸综合征RT-PCR诊断方法的建立

Establishment of A RT-PCR Method for Diagnosis of Porcine Reproductive and Respiratory Syndrome

为建立一种方便快捷的检测猪繁殖与呼吸综合征的RT-PCR方法,根据GenBank中已发表的猪繁殖与呼吸综合征病毒GP5基因设计一对引物,并以合成的cDNA为模板,初步建立了检测猪繁殖与呼吸综合征的RT-PCR方法,进而对对照病毒及采集的病料组织进行了特异性、敏感性和重复性检测。结果显示,该研究建立的RT-PCR方法敏感性高,最低检测阈为10 TCID50/mL,扩增产物的特异性良好,重复性良好,可用于猪繁殖与呼吸综合征的临床诊断及流行病学调查。

In order to develop a RT-PCR method for the rapid and convenient diagnosis of porcine reproductive and respiratory syndrome,a pair of primers were designed according to the porcine reproductive and respiratory syndrome virus gene GP5 published in the GenBank,the synthetical cDNA was used as the template,and a RT-PCR method was established to detect porcine reproductive and respiratory syndrome virus.The specificity,sensitivity and repeatability of the assay were determined by detections of the control virus and the diseased tissue samples.The results indicated that the established RT-PCR method was highly sensitive,and the minimum threshold of detecting the isolated virus was 10 TCID50/mL.The specificity and repeatability of this RT-PCR method were good,and it could be used as an available tool for the clinical diagnosis and epidemiological investigation of porcine reproductive and respiratory syndrome.