National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3M4, Canada.
Viruses. 2024 Sep 10;16(9):1438. doi: 10.3390/v16091438.
Foot-and-mouth disease (FMD) is one of the most infectious viral transboundary diseases of livestock, which causes devastating global economic losses. Different enzyme-linked immunosorbent assays (ELISAs) are used for sero-surveillance of the foot-and-mouth disease virus (FMDV). However, more sensitive, accurate, and convenient ELISAs are still required to detect antibodies against FMDV serotypes. The primary goal of this study was to establish serotype-specific monoclonal antibody (mAb)-based blocking ELISAs (mAb-bELISAs) that would provide better performance characteristics or be equivalent in performance characteristics compared with a conventional polyclonal antibody (pAb)-based competitive ELISA (pAb-cELISA). Four mAb-bELISAs were developed using FMDV serotype-specific mAbs for the detection of anti-FMDV/O/A/Asia1/SAT2 antibodies. Using a 50% cut-off, all four mAb-bELISAs exhibited species-independent 99.74%, 98.01%, 96.59%, and 98.55% diagnostic specificity (DSp) and 98.93%, 98.25%, 100%, and 87.50% diagnostic sensitivity (DSe) for FMDV serotypes O, A, Asia1, and SAT2, respectively. In addition, a 100% DSe of serotypes O- and SAT2-specific mAb-bELISAs was observed for porcine sera when the cut-off was 30%. All mAb-bELISAs developed in this study displayed high repeatability/reproducibility without cross-reactivity. Finally, the diagnostic performance of mAb-bELISAs was found to be better than or equivalent to compared with pAb-cELISAs, suggesting that mAb-bELISAs can be used to replace existing pAb-ELISAs for the detection of antibodies against these four FMDV serotypes.
口蹄疫（FMD）是最具传染性的动物传染病之一，它会给全球经济造成巨大的损失。不同的酶联免疫吸附测定（ELISA）被用于口蹄疫病毒（FMDV）的血清学监测。然而，为了检测针对口蹄疫不同血清型的抗体，仍需要更敏感、更准确和更方便的 ELISA。本研究的主要目的是建立基于单克隆抗体（mAb）的血清型特异性阻断 ELISA（mAb-bELISA），与传统的基于多克隆抗体（pAb）的竞争 ELISA（pAb-cELISA）相比，这些 mAb-bELISA 具有更好的性能特征或具有同等的性能特征。本研究使用 4 种针对 FMDV 不同血清型的特异性 mAb 建立了 4 种 mAb-bELISA，用于检测抗 FMDV/O/A/Asia1/SAT2 抗体。使用 50%的截断值，所有 4 种 mAb-bELISA 对 FMDV 血清型 O、A、Asia1 和 SAT2 的物种独立诊断特异性（DSp）分别为 99.74%、98.01%、96.59%和 98.55%，诊断敏感性（DSe）分别为 98.93%、98.25%、100%和 87.50%。此外，当截断值为 30%时，O 型和 SAT2 型特异性 mAb-bELISA 对猪血清的 DSe 为 100%。本研究中建立的所有 mAb-bELISA 均显示出较高的重复性/再现性，无交叉反应。最后，发现 mAb-bELISA 的诊断性能优于或与 pAb-cELISA 相当，表明 mAb-bELISA 可用于替代现有的 pAb-ELISA 来检测这四种 FMDV 血清型的抗体。
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