Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS, United States.
Foreign Animal Disease Diagnostic Laboratory, National Veterinary Services Laboratories, Animal and Plant Health Inspection Services, United States Department of Agriculture, Plum Island Animal Disease Center, New York, NY, United States.
Virus Res. 2020 Apr 2;279:197871. doi: 10.1016/j.virusres.2020.197871. Epub 2020 Jan 28.
In the absence of a vaccine for African swine fever virus (ASFV), diagnostic tools are critical for early detection and implementation of control measures. Along with other immunogenic proteins, p54 is a good serological target for conducting ASF detection and surveillance. In this study, a panel of 12 mouse monoclonal antibodies (mAbs) was prepared against a baculovirus-expressed p54(60-178) polypeptide. Further screening showed that five mAbs were positive for reactivity against ASFV-infected cells and recombinant p54 proteins. Mapping studies using five polypeptides and 12 oligopeptides, showed that mAb #154-1 recognized a conserved polypeptide sequence, p54(65-75), and was placed into Group 1. Mabs #143-1 and #7 recognized a region covered by p54(93-113) and were placed into Group 2. Group 3 consisted of mAbs #101 and #117, which recognized p54(118-127). Sera from pigs infected with the low virulent OURT 88/3 strain recognized the same p54 region covered by the Group 3 mAbs. When tested in a neutralization format, only mAb #143-1 showed neutralization activity above background. Together, the results identify important antigenic and immunogenic regions located on p54, which provide new tools for improving ASFV diagnostics.
在没有非洲猪瘟病毒 (ASFV) 疫苗的情况下,诊断工具对于早期检测和实施控制措施至关重要。与其他免疫原性蛋白一样,p54 是进行 ASF 检测和监测的良好血清学靶标。在这项研究中,针对杆状病毒表达的 p54(60-178)多肽制备了一组 12 种鼠单克隆抗体 (mAb)。进一步筛选表明,有 5 种 mAb 对感染 ASFV 的细胞和重组 p54 蛋白呈阳性反应。使用 5 种多肽和 12 种寡肽进行的作图研究表明,mAb #154-1 识别一个保守的多肽序列 p54(65-75),并被归入第 1 组。mAbs #143-1 和 #7 识别 p54(93-113)覆盖的区域,被归入第 2 组。第 3 组由 mAbs #101 和 #117 组成,它们识别 p54(118-127)。感染低毒力 OURT 88/3 株的猪血清识别第 3 组 mAb 覆盖的相同 p54 区域。在中和格式中进行测试时,只有 mAb #143-1 显示出高于背景的中和活性。总之,这些结果确定了位于 p54 上的重要抗原和免疫原性区域,为改进 ASFV 诊断提供了新的工具。