Wang Lei, Li Duan, Zeng Daoping, Wang Xiaomin, Liu Yanlin, Peng Guoliang, Xu Zheng, Song Changxu
College of Life Sciences, Longyan University, Longyan 364012, China.
State Key Laboratory of Swine and Poultry Breeding Industry, National Engineering Center for Swine Breeding Industry, College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
Viruses. 2024 Dec 17;16(12):1925. doi: 10.3390/v16121925.
Porcine circovirus 3 (PCV3) is a small non-enveloped circovirus associated with porcine dermatitis and nephropathy syndrome (PDNS). It has occurred worldwide and poses a serious threat to the pig industry. However, there is no commercially available vaccine. PCV3 capsid protein (Cap) is an ideal antigen candidate for serodiagnosis. Here, a novel fully automated chemiluminescence enzyme immunoassay (CLEIA) was developed to detect antibodies (Abs) to Cap in porcine serum. Recombinant PCV3 Cap, self-assembled into virus-like particles (VLPs), was produced using baculovirus and coupled to magnetic particles (Cap-MPs) as carriers. Combined with an alkaline phosphatase (AP)-adamantane (AMPPD) system, Cap-Abs can be rapidly measured on a fully automated chemiluminescence analyzer. Under optimal conditions, a cut-off value of 31,508 was determined, with a diagnostic sensitivity of 96.8% and specificity of 97.3%. No cross-reactivity was observed with PCV1 and PCV2 and other common porcine pathogens, and both intra-assay and inter-assay coefficients were less than 5% and 10%, respectively. Prepared Cap-MPs can be stored at 4 °C for more than 6 months. Importantly, this CLEIA had a good agreement of 95.19% with the commercially available kit, demonstrating excellent analytical sensitivity and significantly reduced operating time and labor. A serological survey was then conducted, and showed that PCV3 continues to spread widely in South China. In conclusion, our CLEIA provides time and labor-saving, and a reliable tool for PCV3 epidemiological surveillance.
猪圆环病毒3型（PCV3）是一种与猪皮炎和肾病综合征（PDNS）相关的小型无包膜圆环病毒。它已在全球范围内出现，对养猪业构成严重威胁。然而，目前尚无商业化疫苗。PCV3衣壳蛋白（Cap）是血清学诊断的理想抗原候选物。在此，开发了一种新型全自动化学发光酶免疫分析方法（CLEIA），用于检测猪血清中针对Cap的抗体（Abs）。使用杆状病毒生产重组PCV3 Cap，其自组装成病毒样颗粒（VLPs），并与磁性颗粒（Cap-MPs）偶联作为载体。结合碱性磷酸酶（AP）-金刚烷（AMPPD）系统，可在全自动化学发光分析仪上快速检测Cap-Abs。在最佳条件下，确定的临界值为31,508，诊断敏感性为96.8%，特异性为97.3%。未观察到与PCV1和PCV2以及其他常见猪病原体的交叉反应，批内和批间系数分别小于5%和10%。制备的Cap-MPs可在4℃下储存6个月以上。重要的是，该CLEIA与市售试剂盒的一致性良好，为95.19%，显示出优异的分析灵敏度，显著减少了操作时间和劳动力。随后进行了血清学调查，结果表明PCV3在中国南方继续广泛传播。总之，我们的CLEIA为PCV3流行病学监测提供了省时省力且可靠的工具。
