基于重组NS1蛋白猪细小病毒野毒抗体间接ELISA诊断方法的建立与应用

本研究以原核表达的重组NS1蛋白作为诊断抗原,建立了检测猪细小病毒(PPV)野毒抗体的NS1-ELISA诊断方法。该方法检测猪瘟病毒、猪繁殖与呼吸综合征病毒、猪伪狂犬病病毒、猪圆环病毒2型、猪流行性腹泻病毒5种常见猪病病毒的阳性血清均为阴性;检测灵敏度为1:12800;批内、批间重复性试验的变异系数分别小于5%和10%;与血凝抑制试验(HI)符合率为100%。本研究建立的PPV NS1-ELISA检测方法具有良好的特异性、敏感性和重复性,为PPV的野毒抗体检测及PPV流行病学调查等快速诊断提供了一种技术手段。

Establishment and Application of an Indirect ELISA for Detecting Wild Virus Antibodies to Porcine Parvovirus using NS1 Recombinant Protein

Using recombinant NS1 protein as coating antigen, an indirect ELISA was successfully developed to detect wild virus antibody to PPV. The results of detecting the positive sera of other five swine diseases(CSFV, PRRSV, PRV, PCV2, PEDV) using the method were negative. Coefficient of variability percent (C.V%) of intro-batch and inter-batch duplicative tests was less than 5% and 10%, respectively. Lowest antibody titer of positive control serum was 1:12800. Comparing with the HI assay, the concordance rate was 100%. Therefore, the NS1-ELISA had good specificity, sensitivity and repeatability, and could be used as a simple and rapid serology detection method for monitoring the anti-PPV wild virus antibody and epidemiologic survey of PPV.