State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China.
Regeneron Pharmaceuticals Inc., 777 Old Saw Mill River Road, Tarrytown, New York, NY 10591, USA.
Viruses. 2022 Aug 20;14(8):1828. doi: 10.3390/v14081828.
Porcine parvovirus (PPV) is widely prevalent in pig farms. PPV is closely related to porcine respiratory disease complex (PRDC) and porcine circovirus disease (PCVD), which seriously threatens the healthy development of the pig industry. Although commercial antibody detection kits are available, they are expensive and unsuitable for large-scale clinical practice. Here, a soluble VP2 protein of PPV is efficiently expressed in the expression system. The VP2 protein can be self-assembled into virus-like particles (VLPs) in vitro. After multiple steps of chromatography purification, PPV-VLPs with a purity of about 95% were obtained. An indirect, enzyme-linked immunosorbent assay (I-ELISA), comparable to a commercial PPV kit, was developed based on the purified PPV-VLPs and was used to detect 487 clinical pig serum samples. The results showed that the I-ELISA is a simple, cost-effective, and efficient method for the diagnosis of clinical pig serum and plasma samples. In summary, high-purity, tag-free PPV-VLPs were prepared, and the established VLP-based I-ELISA is of great significance for the sero-monitoring of antibodies against PPV.
猪细小病毒（PPV）在养猪场中广泛流行。PPV 与猪呼吸道疾病复合症（PRDC）和猪圆环病毒病（PCVD）密切相关，严重威胁着养猪业的健康发展。虽然有商品化的抗体检测试剂盒，但价格昂贵，不适合大规模临床实践。本研究在表达系统中高效表达了 PPV 的可溶性 VP2 蛋白。该 VP2 蛋白可在体外自行组装成病毒样颗粒（VLPs）。经过多次层析纯化步骤，获得了纯度约为 95%的 PPV-VLPs。基于纯化的 PPV-VLPs 建立了间接酶联免疫吸附试验（I-ELISA），与商品化的 PPV 试剂盒相当，并用于检测 487 份临床猪血清样本。结果表明，I-ELISA 是一种简单、经济高效的用于诊断临床猪血清和血浆样本的方法。综上所述，制备了高纯度、无标签的 PPV-VLPs，并建立了基于 VLP 的 I-ELISA，这对于 PPV 抗体的血清学监测具有重要意义。
