Wang Lei, Li Duan, Liu Yanlin, Zhang Leyi, Peng Guoliang, Xu Zheng, Jia Hong, Song Changxu
College of Animal Science and National Engineering Center for Swine Breeding Industry, South China Agricultural University, Guangzhou, China.
Henry Fok School of Biology and Agriculture, Shaoguan University, Shaoguan, China.
Front Vet Sci. 2023 Jun 9;10:1160583. doi: 10.3389/fvets.2023.1160583. eCollection 2023.
African swine fever (ASF), caused by ASF virus (ASFV), is a highly contagious and lethal disease of domestic pigs leading to tremendous economic losses. As there are no vaccines and drugs available. An effective diagnosis to eliminate ASFV-infected pigs is a crucial strategy to prevent and control ASF. To this end, ASFV capsid protein p72 was expressed using Chinese hamster ovary (CHO) cells and subsequently conjugated with horseradish peroxidase (HRP) to develop a one-step double-antigen sandwich enzyme-linked immunosorbent assay (one-step DAgS-ELISA). The performance of this ELISA for detecting ASFV antibodies was evaluated. Overall, a diagnostic sensitivity of 97.96% and specificity of 98.96% was achieved when the cutoff value was set to 0.25. No cross-reaction with healthy pig serum and other swine viruses was observed. The coefficients of variation of the intra-assay and inter-assay were both <10%. Importantly, this ELISA could detect antibodies in standard serum with 12,800-fold dilution, and seroconversion started from the 7th day post-inoculation (dpi), showing excellent analytical sensitivity and great utility. Furthermore, compared to the commercial kit, this ELISA had a good agreement and significantly shorter operation time. Collectively, a novel one-step DAgS-ELISA for detecting antibodies against ASFV is developed, which will be reliable and convenient to monitor ASFV infection.
非洲猪瘟（ASF）由非洲猪瘟病毒（ASFV）引起，是家猪的一种高度传染性和致死性疾病，会导致巨大的经济损失。由于目前尚无可用的疫苗和药物，有效诊断以淘汰感染ASFV的猪是预防和控制ASF的关键策略。为此，利用中国仓鼠卵巢（CHO）细胞表达了ASFV衣壳蛋白p72，随后将其与辣根过氧化物酶（HRP）偶联，开发了一种一步双抗原夹心酶联免疫吸附测定法（一步DAgS-ELISA）。评估了该ELISA检测ASFV抗体的性能。总体而言，当临界值设定为0.25时，诊断敏感性达到97.96%，特异性达到98.96%。未观察到与健康猪血清和其他猪病毒的交叉反应。批内和批间变异系数均<10%。重要的是，该ELISA能够检测稀释12,800倍的标准血清中的抗体，并且在接种后第7天（dpi）开始出现血清转化，显示出优异的分析敏感性和实用性。此外，与商业试剂盒相比，该ELISA具有良好的一致性且操作时间显著缩短。总体而言，开发了一种用于检测抗ASFV抗体的新型一步DAgS-ELISA，这将为监测ASFV感染提供可靠且便捷的方法。
