Beijing Kemufeng Biopharmaceutical Co., Ltd, No.25 Xiangrui Street Daxing District, Beijing, 102600, China.
BMC Vet Res. 2023 Jul 19;19(1):87. doi: 10.1186/s12917-023-03648-7.
Porcine circovirus 2 (PCV-2) is one of the pathogens that leads to a growing and persistent threat in pigs. Thus, the development of serological detection methods for PCV-2 is of great necessity for clinical diagnosis as well as epidemiological investigations. This study aimed to establish an indirect enzyme-linked immunosorbent assay (ELISA) to examine antibodies against PCV-2 based on virus-like particles (VLPs).
Recombinant PCV-2 Cap protein was expressed in the baculovirus-insect cells system and PCV-2 VLPs were observed over transmission electron microscopy (TEM). The PCV-2 VLPs were shown to have good immunogenicity in mice and stimulated a high level of PCV-2 antibody titers. Using PCV-2 VLPs as coating antigen, the indirect ELISA can detect PCV-2 antibodies in animals with diagnostic sensitivity and specificity of 98.33% and 93.33% compared to immunofluorescence assay (IFA), respectively. The intra- and inter-assay coefficient variations (CVs) were < 10% in a batch, and < 15% in different batches, indicating good repeatability. There was no cross-reaction of this ELISA with antibodies against other porcine viruses. A total of 170 serum samples collected from different pig farms in China were tested for PCV-2 antibodies, and 151 (88.8%) samples were PCV-2 antibody positive.
Our findings suggest that this ELISA was rapid, specific, and reproducible and can be used for large-scale serological investigations of PCV-2 antibodies in pigs.
猪圆环病毒 2 型（PCV-2）是导致猪群持续增长的威胁之一。因此，开发用于检测 PCV-2 的血清学检测方法对于临床诊断和流行病学调查非常必要。本研究旨在建立一种基于病毒样颗粒（VLPs）的间接酶联免疫吸附试验（ELISA）来检测针对 PCV-2 的抗体。
重组 PCV-2 Cap 蛋白在杆状病毒-昆虫细胞系统中表达，并通过透射电子显微镜（TEM）观察到 PCV-2 VLPs。PCV-2 VLPs 在小鼠中表现出良好的免疫原性，并刺激产生高水平的 PCV-2 抗体滴度。使用 PCV-2 VLPs 作为包被抗原，间接 ELISA 可检测动物中的 PCV-2 抗体，与免疫荧光分析（IFA）相比，其诊断敏感性和特异性分别为 98.33%和 93.33%。批内和批间变异系数（CV）均<10%，不同批次间<15%，表明重复性良好。该 ELISA 与针对其他猪病毒的抗体无交叉反应。共检测了来自中国不同猪场的 170 份血清样本，其中 151 份（88.8%）样本为 PCV-2 抗体阳性。
本研究表明，该 ELISA 快速、特异、可重复，可用于大规模检测猪 PCV-2 抗体。
