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检测牛针对保守衣壳表位的抗体,作为口蹄疫新型通用血清学检测方法的基础。

Detection of Bovine Antibodies against a Conserved Capsid Epitope as the Basis of a Novel Universal Serological Test for Foot-and-Mouth Disease.

机构信息

The Pirbright Institute, Woking, United Kingdom

The Pirbright Institute, Woking, United Kingdom.

出版信息

J Clin Microbiol. 2020 May 26;58(6). doi: 10.1128/JCM.01527-19.

Abstract

Diagnostic tests for foot-and-mouth disease (FMD) include the detection of antibodies against either the viral nonstructural proteins or the capsid. The detection of antibodies against the structural proteins (SP) of the capsid can be used to monitor seroconversion in both infected and vaccinated animals. However, SP tests need to be tailored to the individual FMD virus (FMDV) serotype and their sensitivity may be affected by antigenic variability within each serotype and mismatching between test reagents. As a consequence, FMD reference laboratories are required to maintain multiple type-specific SP assays and reagents. A universal SP test would simplify frontline diagnostics and facilitate large-scale serological surveillance and postvaccination monitoring. In this study, a highly conserved region in the N terminus of FMDV capsid protein VP2 (VP2N) was characterized using a panel of intertype-reactive monoclonal antibodies. This revealed a universal epitope in VP2N which could be used as a peptide antigen to detect FMDV-specific antibodies against all types of the virus. A VP2-peptide enzyme-linked immunosorbent assay (VP2-ELISA) was optimized using experimental and reference antisera from immunized, convalescent, and naïve animals ( = 172). The VP2-ELISA is universal and simple and provided sensitive (99%) and specific (93%) detection of antibodies to all FMDV strains used in this study. We anticipate that this SP test could have utility for serosurveillance during virus incursions in FMD-free countries and as an additional screening tool to assess FMD virus circulation in countries where the disease is endemic.

摘要

口蹄疫(FMD)的诊断检测包括针对病毒非结构蛋白或衣壳的抗体检测。针对衣壳结构蛋白(SP)的抗体检测可用于监测感染和接种动物的血清转化。然而,SP 检测需要针对特定的 FMD 病毒(FMDV)血清型进行定制,其敏感性可能受到每个血清型内的抗原变异性和检测试剂之间的不匹配的影响。因此,FMD 参考实验室需要维护多种针对特定类型的 SP 检测方法和试剂。通用 SP 检测将简化一线诊断,并促进大规模血清学监测和接种后监测。在这项研究中,使用一组交叉反应性单克隆抗体对 FMDV 衣壳蛋白 VP2(VP2N)的 N 端高度保守区进行了表征。这揭示了 VP2N 中的一个通用表位,可用作肽抗原来检测针对所有类型病毒的 FMDV 特异性抗体。使用来自免疫、康复和未感染动物的实验和参考血清( = 172)优化了 VP2-肽酶联免疫吸附试验(VP2-ELISA)。VP2-ELISA 是通用且简单的,能够敏感(99%)和特异性(93%)地检测本研究中使用的所有 FMDV 毒株的抗体。我们预计,这种 SP 检测方法可用于在无口蹄疫国家发生病毒入侵期间进行血清学监测,并作为评估疾病流行地区 FMD 病毒传播的额外筛选工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1af7/7269384/fc1778fe6560/JCM.01527-19-f0001.jpg

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