猪细小病毒3型VP2蛋白主要抗原表位区的原核表达及间接ELISA方法的建立

Establishment of an indirect ELISA assay of porcine parvovirus 3 with the major epitope domain of VP2 as coating antigen

为建立检测猪细小病毒3型(PPV3)的间接ELISA方法,本研究选取PPV3 VP2多表位亲水区序列设计引物,扩增并原核表达该基因片段,以表达的重组蛋白作为包被抗原建立了PPV3间接ELISA抗体检测方法。结果显示,该检测方法仅对PPV3血清检测为阳性,与PPV1、PPV2、猪瘟病毒、口蹄疫病毒、猪圆环病毒2型、猪伪狂犬病毒和猪繁殖障碍与呼吸综合征病毒等的标准阳性血清均无交叉反应,具有较强的特异性;批内和批间重复性试验的变异系数均小于10%,具有良好的重复性。本研究为临床上PPV3的检测提供了血清学检测手段。

To establish a method for detecting porcine parvovirus 3 （PPV3）, an indirect ELISA was developed with the recombinant protein as coating antigen, which was expressed from partial VP2 gene encoding multi-epitope and strong hydrophilia of PPV3. This method possessed high specificity for detection of antibody against PPV3, but had no cross-reactions with the positive serum, such as classical swine fever virus, foot and mouth disease virus, porcine circovirus type 2, pseudorabies virus, porcine reproductive and respiratory syndrome virus, PPV1 and PPV2. The variability of intra- and inter-batch repeatability were under 10%. This method might be served as an efficient serological assay for the detection of PPV3 in clinic.