Foreign Animal Disease Division, Animal and Plant Quarantine Agency, Gimcheon, Gyeongsangbuk-do 39660, the Republic of Korea.
Median Diagnostics, Chuncheon, Gangwon-do 24399, the Republic of Korea.
J Virol Methods. 2022 Nov;309:114611. doi: 10.1016/j.jviromet.2022.114611. Epub 2022 Sep 2.
African swine fever (ASF) is a highly fatal viral disease affecting pigs. It is caused by the ASF virus (ASFV), and causes serious economic losses to the swine industry worldwide, including in Korea. Commercially available enzyme-linked immunosorbent assay (ELISA) kits for detecting anti-ASFV antibodies are used for the diagnosis and surveillance of ASF. In this study, an ELISA was developed to detect anti-ASFV antibodies using two recombinant proteins, p22 and p30, from genotype II ASFV. Recombinant transmembrane domain-deleted p22 (p22∆TM) and p30 were expressed in E.coli vector system pET32a and mixed for use as antigens in indirect ELISA. The p22∆TM/p30-based indirect ELISA was validated using 31 sera from genotype I ASFV-infected pigs and 1133 sera from uninfected pigs. Area under the curve of this test was 0.999 [95 % concentration interval 0.992-1.000], and sensitivity and specificity were 93.5 % and 99.8 %, respectively. The between run coefficient of variation for internal quality control serum was 6.61 %. In the seroconversion analysis, the p22∆TM/p30-based indirect ELISA showed equal or better ability to detect antibodies in pigs experimentally challenged with ASFV p72 genotypes I and II (p < 0.05). In conclusion, the p22∆TM/p30-based indirect ELISA is a reliable diagnostic method for detecting anti-ASFV antibodies.
非洲猪瘟 (ASF) 是一种严重危害猪只的高致命性病毒性疾病。它由 ASF 病毒 (ASFV) 引起，给全球的养猪业造成了严重的经济损失，包括韩国。市售的用于检测抗 ASFV 抗体的酶联免疫吸附试验 (ELISA) 试剂盒用于 ASF 的诊断和监测。在这项研究中，我们使用来自基因型 II ASFV 的两个重组蛋白 p22 和 p30 开发了一种用于检测抗 ASFV 抗体的 ELISA。在大肠杆菌表达载体系统 pET32a 中表达了重组跨膜结构域缺失的 p22 (p22∆TM) 和 p30，并将它们混合用作间接 ELISA 的抗原。p22∆TM/p30 基于间接 ELISA 用来自基因型 I ASFV 感染猪的 31 份血清和 1133 份未感染猪的血清进行了验证。该试验的曲线下面积为 0.999 [95 %置信区间 0.992-1.000]，灵敏度和特异性分别为 93.5 %和 99.8 %。内部质量控制血清的批间变异系数为 6.61 %。在血清转化分析中，p22∆TM/p30 基于间接 ELISA 在检测实验性感染 ASFV p72 基因型 I 和 II 的猪的抗体方面显示出同等或更好的能力 (p < 0.05)。总之，p22∆TM/p30 基于间接 ELISA 是一种可靠的检测抗 ASFV 抗体的诊断方法。
