State Key Laboratory for Animal Disease Control and Prevention, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
Gansu Province Research Center for Basic Disciplines of Pathogen Biology, Lanzhou Veterinary Research Institute Chinese Academy of Agricultural Sciences, Lanzhou, China.
Appl Microbiol Biotechnol. 2024 Aug 2;108(1):431. doi: 10.1007/s00253-024-13249-5.
African swine fever (ASF) is a highly contagious and fatal viral disease that has caused huge economic losses to the pig and related industries worldwide. At present, rapid, accurate, and sensitive laboratory detection technologies are important means of preventing and controlling ASF. However, because attenuated strains of African swine fever virus (ASFV) are constantly emerging, an ASFV antibody could be used more effectively to investigate the virus and control the disease on pig farms. The isolation of ASFV-specific antibodies is also essential for the diagnosis of ASF. Therefore, in this study, we developed two chemiluminescence immunoassays (CLIAs) to detect antibodies directed against ASFV p72: a traditional plate-type blocking CLIA (p72-CLIA) and an automatic tubular competitive CLIA based on magnetic particles (p72-MPCLIA). We compared the diagnostic performance of these two methods to provide a feasible new method for the effective prevention and control of ASF and the purification of ASFV. The cut-off value, diagnostic sensitivity (Dsn), and diagnostic specificity (Dsp) of p72-CLIA were 40%, 100%, and 99.6%, respectively, in known background serum, whereas those of p72-MPCLIA were 36%, 100%, and 99.6%, respectively. Thus, both methods show good Dsn, Dsp, and repeatability. However, when analytical sensitivity was evaluated, p72-MPCLIA was more sensitive than p72-CLIA or a commercial enzyme-linked immunosorbent assay. More importantly, p72-MPCLIA reduced the detection time to 15 min and allowed fully automated detection. In summary, p72-MPCLIA showed superior diagnostic performance and offered a new tool for detecting ASFV infections in the future. KEY POINTS: • Two chemiluminescence immunoassay (plate-type CLIA and tubular CLIA) methods based on p72 monoclonal antibody (mAb) were developed to detect ASFV antibody. • Both methods show good diagnostic performance (Dsn (100%), Dsp (99.6%), and good repeatability), and p72-MPCLIA detects antibodies against ASFV p72 with high efficiency in just 15 min.
非洲猪瘟（ASF）是一种高度传染性和致命性的病毒性疾病，已在全球范围内给猪和相关产业造成了巨大的经济损失。目前，快速、准确、灵敏的实验室检测技术是预防和控制 ASF 的重要手段。然而，由于非洲猪瘟病毒（ASFV）的减毒株不断出现，ASFV 抗体的使用可以更有效地调查病毒并控制猪场的疾病。ASF 特异性抗体的分离对于 ASF 的诊断也是必不可少的。因此，在这项研究中，我们开发了两种检测针对 ASFV p72 的抗体的化学发光免疫分析（CLIAs）：传统平板型阻断 CLIA（p72-CLIA）和基于磁性颗粒的自动管状竞争 CLIA（p72-MPCLIA）。我们比较了这两种方法的诊断性能，为 ASF 的有效预防和控制以及 ASFV 的纯化提供了一种可行的新方法。在已知背景血清中，p72-CLIA 的截断值、诊断灵敏度（Dsn）和诊断特异性（Dsp）分别为 40%、100%和 99.6%，而 p72-MPCLIA 的分别为 36%、100%和 99.6%。因此，两种方法均具有良好的 Dsn、Dsp 和重复性。然而，当评估分析灵敏度时，p72-MPCLIA 比 p72-CLIA 或商业酶联免疫吸附测定更灵敏。更重要的是，p72-MPCLIA 将检测时间缩短至 15 分钟，并允许全自动检测。综上所述，p72-MPCLIA 显示出优越的诊断性能，为未来检测 ASFV 感染提供了新工具。
• 基于 p72 单克隆抗体（mAb）开发了两种化学发光免疫分析（平板型 CLIA 和管状 CLIA）方法来检测 ASFV 抗体。
• 两种方法均具有良好的诊断性能（Dsn（100%）、Dsp（99.6%）和良好的重复性），p72-MPCLIA 在 15 分钟内即可高效检测针对 ASFV p72 的抗体。
