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A recombinant nucleocapsid protein-based indirect enzyme-linked immunosorbentassay to detect antibodies against porcine deltacoronavirus

机译:基于重组核衣壳蛋白的间接酶联免疫吸附剂法检测针对猪三角洲冠状病毒的抗体

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摘要

Recently, porcine deltacoronavirus (PDCoV) has been proven to be associated with enteric disease in piglets. Diagnostic tools for serological surveys of PDCoV remain in the developmental stage when compared with those for other porcine coronaviruses. In our study, an indirect enzyme-linked immunosorbent assay (ELISA) (rPDCoV-N-ELISA) was developed to detect antibodies against PDCoV using a histidine-tagged recombinant nucleocapsid (N) protein as an antigen. The rPDCoV-N-ELISA did not cross-react with antisera against porcine epidemic diarrhea virus, swine transmissible gastroenteritis virus, porcine group A rotavirus, classical swine fever virus, porcine circovirus-2, porcine pseudorabies virus, and porcine reproductive and respiratory syndrome virus; the receiver operating characteristic (ROC) curve analysis revealed 100% sensitivity and 90.4% specificity of the rPDCoV-N-ELISA based on samples of known status (n=62). Analyses of field samples (n=319) using the rPDCoV-N-ELISA indicated that 11.59% of samples were positive for antibodies against PDCoV. These data demonstrated that the rPDCoV-N-ELISA can be used for epidemiological investigations of PDCoV and that PDCoV had a low serum prevalence in pig population in Heilongjiang province, northeast China.
机译:最近,猪三角冠状病毒(PDCoV)已被证明与仔猪的肠道疾病有关。与其他猪冠状病毒相比,PDCoV血清学检查的诊断工具仍处于开发阶段。在我们的研究中,开发了一种间接酶联免疫吸附测定(ELISA)(rPDCoV-N-ELISA),以组氨酸标记的重组核衣壳(N)蛋白作为抗原来检测针对PDCoV的抗体。 rPDCoV-N-ELISA不能与针对猪流行性腹泻病毒,猪可传播的肠胃炎病毒,猪A型轮状病毒,经典猪瘟病毒,猪圆环病毒2,猪伪狂犬病毒以及猪繁殖与呼吸综合征病毒的抗血清交叉反应;接收者工作特征(ROC)曲线分析显示,基于已知状态的样品(n = 62),rPDCoV-N-ELISA的敏感性为100%,特异性为90.4%。使用rPDCoV-N-ELISA对现场样品(n = 319)进行的分析表明,有11.59%的样品对PDCoV的抗体呈阳性。这些数据表明,rPDCoV-N-ELISA可以用于PDCoV的流行病学研究,并且PDCoV在中国东北黑龙江省的猪群中的血清患病率较低。

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