Acta Virol. 2021;65(3):273-278. doi: 10.4149/av_2021_304.
African swine fever (ASF) is an acute and severe infectious disease that seriously endangers the global porcine industry. In order to develop ASF serodiagnostic reagents with high specificity and sensitivity, in the present study, the antigenic epitopes of P72 protein of African swine fever virus (ASFV) were analyzed, and the ASFV multi-epitope fusion gene MeP72 in tandem with the dominant linear epitopes was constructed. The recombinant multi-epitope fusion MeP72 (reMeP72) was prepared in Escherichia coli. A colloidal gold-based immunochromatographic assay (CGIA) based on reMeP72 was developed for the detection of antibodies against ASFV. A total of 139 pig clinical serum samples were used for assessment of the potential diagnostic value of reMeP72. The results showed that CGIA did not cross-react with positive sera of viruses, such as classical swine fever virus (CSFV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV) and swine influenza virus (SIV), showing high specificity. Sensitivity analysis showed that CGIA could detect ASFV-positive serum at a dilution of 1:64. Compared with commercial ASFV kits, the sensitivity and specificity of ASFV CGIA based on reMeP72 protein were 85.7% and 97.6%, respectively. The agreement rate of the two methods was 96.4%, showing a good detection performance. The results indicated that the reMeP72 was of potential value for the serodiagnosis of ASF. Keywords: African swine fever virus; P72 gene; antigenic protein; colloidal gold-based immunochromatographic assay.
非洲猪瘟（ASF）是一种严重危害全球养猪业的急性、烈性传染病。为了研发具有高特异性和高灵敏度的 ASF 血清学诊断试剂，本研究对非洲猪瘟病毒（ASFV）P72 蛋白的抗原表位进行分析，并串联 ASFV 优势线性表位构建 ASFV 多表位融合基因 MeP72。在大肠杆菌中制备了重组多表位融合 MeP72（reMeP72）。以 reMeP72 为基础建立了胶体金免疫层析法（CGIA），用于检测抗 ASFV 抗体。用 139 份猪临床血清样本评估 reMeP72 的潜在诊断价值。结果显示，CGIA 与猪瘟病毒（CSFV）、猪圆环病毒 2 型（PCV2）、猪伪狂犬病病毒（PRV）、猪繁殖与呼吸综合征病毒（PRRSV）和猪流感病毒（SIV）等阳性血清无交叉反应，具有很高的特异性。敏感性分析表明，CGIA 可检测到稀释度为 1:64 的 ASFV 阳性血清。与商业 ASFV 试剂盒相比，基于 reMeP72 蛋白的 ASFV CGIA 的灵敏度和特异性分别为 85.7%和 97.6%。两种方法的符合率为 96.4%，检测性能良好。结果表明，reMeP72 对 ASF 的血清学诊断具有潜在价值。关键词：非洲猪瘟病毒；P72 基因；抗原蛋白；胶体金免疫层析法。
