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基于单克隆抗体的固相竞争 ELISA 检测口蹄疫病毒 A 型特异性抗体。

Monoclonal antibody based solid phase competition ELISA to detect FMDV serotype A specific antibodies.

机构信息

FMD QC laboratory, ICAR-Indian Veterinary Research Institute, H A Farm, Hebbal, Bengaluru, Karnataka 560 024, India.

FMD QC laboratory, ICAR-Indian Veterinary Research Institute, H A Farm, Hebbal, Bengaluru, Karnataka 560 024, India.

出版信息

J Virol Methods. 2024 Jul;328:114959. doi: 10.1016/j.jviromet.2024.114959. Epub 2024 May 23.

Abstract

In Foot-and-mouth disease (FMD) enzootic countries, periodic vaccination is the key tool in controlling the disease incidence. Active seromonitoring of the vaccinated population is critical to assess the impact of vaccination. Virus neutralization test (VNT) and enzyme-linked immunosorbent assays (ELISA) are commonly used for antibody detection. Assays like liquid phase blocking ELISA (LPBE) or solid phase competition ELISA (SPCE) are preferred as they do not require handling of live FMDV and are routinely used for seromonitoring or for vaccine potency testing; however, false positives are high in LPBE. Here we report, a monoclonal antibody (mAb) based SPCE as a potential alternate assay for antibody titration. From a panel of 12 mAbs against FMDV serotype A, two mAbs were chosen for the development of SPCE. Based on a set of 453 sera, it was demonstrated that mAb 2C4G11, mAb 6E8D11and polyclonal antibody (pAb) based SPCE had a relative sensitivity of 86.1, 86.1 and 80.3 %; and specificity of 99.6, 99.1 and 99.1 %, respectively. The correlation, repeatability, and level of agreement of the assays were high demonstrating the potential use of mAb in large scale surveillance studies and regular vaccine potency testing.

摘要

在口蹄疫(FMD)流行国家,定期接种疫苗是控制疾病发病率的关键工具。对接种人群进行主动血清监测对于评估疫苗接种效果至关重要。病毒中和试验(VNT)和酶联免疫吸附试验(ELISA)常用于抗体检测。液相阻断 ELISA(LPBE)或固相竞争 ELISA(SPCE)等检测方法因其不需要处理活 FMDV 而被优先选用,并且常用于血清监测或疫苗效力测试;然而,LPBE 的假阳性率较高。在这里,我们报告了一种基于单克隆抗体(mAb)的 SPCE,作为抗体滴定的潜在替代检测方法。在针对 FMDV 血清型 A 的 12 种 mAb 中,选择了两种 mAb 用于开发 SPCE。基于 453 份血清的研究结果表明,mAb 2C4G11、mAb 6E8D11 和基于多克隆抗体(pAb)的 SPCE 的相对灵敏度分别为 86.1%、86.1%和 80.3%;特异性分别为 99.6%、99.1%和 99.1%。这些检测方法的相关性、重复性和一致性水平都很高,表明 mAb 在大规模监测研究和常规疫苗效力测试中具有潜在的应用价值。

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