African swine fever (ASF) is one of the most complex and lethally haemorrhagic viral diseases of swine, affecting all breeds and ages of pigs. In the absence of ASF vaccines, reliable laboratory diagnosis and restricted biosecurity are critical for disease prevention and control. A detection of ASF‐specific antibodies in an unvaccinated pig is a good marker for the diagnosis of ASF. The immunoperoxidase test (IPT) is a sensitive test for detecting ASF virus (ASFV) antibodies. However, due to the complexity of the procedure, the IPT is only suitable to be used as a confirmatory test. The ASFV p30 protein‐based enzyme‐linked immunosorbent assay (ELISA) is widely used for ASFV antibody screening, but the sensitivity is not comparable to the IPT. It is essential to have a better understanding of the antigenic properties of ASFV p30 to improve p30‐based serologic tests. In this study, we developed a panel of 21 monoclonal antibodies (mAbs) against ASFV p30. With 14 out of the 21 mAbs, we defined 4 antigenic regions that contain at least 4 linear epitopes. Nine of the 14 mAbs mapped to antigenic regions 3 and 4 reacted with p30 in all serologic methods tested in this study, such as indirect immunofluorescence assay (IFA), ELISA and Western blot. The antigenic regions 3 and 4 are highly conserved and immunodominant in host antibody response. These mAbs and the defined p30 antigenic regions 3 and 4 provide valuable tools for the development and improvement of ASF serologic assays.

中文翻译：

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非洲猪瘟病毒磷蛋白P30抗原区

非洲猪瘟（ASF）是猪最复杂、最致命的出血性病毒性疾病之一，影响所有品种和年龄的猪。在缺乏非洲猪瘟疫苗的情况下，可靠的实验室诊断和有限的生物安全对于疾病预防和控制至关重要。在未接种疫苗的猪中检测到 ASF 特异性抗体是诊断 ASF 的良好标志。免疫过氧化物酶试验 (IPT) 是检测非洲猪瘟病毒 (ASFV) 抗体的敏感试验。然而，由于程序的复杂性，IPT仅适合用作验证性测试。基于 ASFV p30 蛋白的酶联免疫吸附测定 (ELISA) 广泛用于 ASFV 抗体筛查，但其灵敏度无法与 IPT 相比。更好地了解 ASFV p30 的抗原特性对于改进基于 p30 的血清学检测至关重要。在这项研究中，我们开发了一组 21 种针对 ASFV p30 的单克隆抗体 (mAb)。对于 21 种 mAb 中的 14 种，我们定义了 4 个包含至少 4 个线性表位的抗原区域。在本研究测试的所有血清学方法中，例如间接免疫荧光测定 (IFA)、ELISA 和蛋白质印迹，映射到抗原区域 3 和 4 的 14 种 mAb 中有 9 种与 p30 发生反应。抗原区3和4是高度保守的并且在宿主抗体反应中具有免疫显性。这些 mAb 和确定的 p30 抗原区域 3 和 4 为 ASF 血清学检测的开发和改进提供了宝贵的工具。