猪Kobu病毒巢式RT-PCR检测方法的建立及初步应用

为了对猪Kobu病毒进行快速诊断及流行病学监测,根据GenBank中公布的9条猪Kobu病毒基因组全序列,在3CD非结构蛋白保守区域设计并合成了2对特异性引物,建立了猪Kobu病毒的巢式RT-PCR检测方法,并用该方法对猪Kobu病毒DX株动物回归试验发病的25头仔猪的组织病料以及送检的72份临床样本进行检测.结果表明,使用该方法对25头发病猪内脏病料的阳性检出率为100％；对送检的72份临床样品的阳性检出率为87.5％ (63/72).该方法特异性强、灵敏度高、重复性好,适用于猪Kobu病毒的临床快速诊断及流行病学监测.

Development and Application of a Nested RT-PCR Assay for Detection of Porcine Kobuvirus

In order to establish a nested RT-PCR assay for diagnosis of porcine kobuviruses, two pairs of specific primers were designed and synthesized in the conserved region of 3CD non-structural proteins according to 9 porcine kobuviruses complete genome in GenBank. The assay was used to detect samples of 25 infected piglets with strain PKV-DX and 72 clinical samples from areas of Beijing, Xinjiang, and Inner Mongolia. The positive rate of 25 samples was 100%, and that of 72 clinical samples was 87.5%. The nested RT-PCR assay with high specificity, sensitivity and repeatability can be applied to the clinical rapid detection and epidemiological surveillance.