口蹄疫病毒(FMDV)的多种血清型和拓扑类型在流行地区流行,在当地造成相当大的影响。此外,引入新领域非常令人关注。的确,近年来,多次FMDV爆发,由从原始区域逃脱的拓扑类型引起的,在世界各地都有记录。在这两种情况下,快速准确的诊断,包括确定引起特定爆发的血清型和拓扑类型,在实施最有效,最适当的措施来控制疾病的传播中起着重要作用。在本研究中,我们描述了在坦桑尼亚北部收集的一组囊泡样本中FMDV的一系列诊断和分型工具的性能(东非,EA)在2012-2018年期间。具体来说,我们用实时RT-PCR靶向3D序列测试了这些样品,用于泛FMDV检测;基于FMDV单克隆抗体的抗原(Ag)检测和血清分型ELISA试剂盒;LFBKαVβ6细胞系上的病毒分离(VI);和一组四个拓扑类型特异性实时RT-PCR,专门为EA中的循环菌株定制。3D实时RT-PCR显示出最高的诊断灵敏度,但缺乏打字能力。Ag-ELISA在71%的样品匀浆中检测并分型了FMDV,而VI联合Ag-ELISA分型的有效率为82%。拓扑类型特异性实时RT-PCR小组在93%的样品中鉴定并分型FMDV。然而,SAT1实时RT-PCR的失败率最高(20%)。简而言之,拓扑类型特异性实时RT-PCR对EAFMDV具有最高的血清分型能力,尽管需要四次检测,而Ag-ELISA,不太敏感,是最方便用户的,因此适用于任何实验室水平。总之,当四个比较测试结合使用时,诊断和血清分型表现均接近100%.
Multiple serotypes and topotypes of foot-and-mouth disease virus (FMDV) circulate in endemic areas, posing considerable impacts locally. In addition, introductions into new areas are of great concern. Indeed, in recent years, multiple FMDV outbreaks, caused by topotypes that have escaped from their original areas, have been recorded in various parts of the world. In both cases, rapid and accurate diagnosis, including the identification of the serotype and topotype causing the given outbreaks, plays an important role in the implementation of the most effective and appropriate measures to control the spread of the disease. In the present study, we describe the performance of a range of diagnostic and typing tools for FMDV on a panel of vesicular samples collected in northern Tanzania (East Africa, EA) during 2012-2018. Specifically, we tested these samples with a real-time RT-PCR targeting 3D sequence for pan-FMDV detection; an FMDV monoclonal antibody-based antigen (Ag) detection and serotyping ELISA kit; virus isolation (VI) on LFBKαVβ6 cell line; and a panel of four topotype-specific real-time RT-PCRs, specifically tailored for circulating strains in EA. The 3D real-time RT-PCR showed the highest diagnostic sensitivity, but it lacked typing capacity. Ag-ELISA detected and typed FMDV in 71% of sample homogenates, while VI combined with Ag-ELISA for typing showed an efficiency of 82%. The panel of topotype-specific real-time RT-PCRs identified and typed FMDV in 93% of samples. However, the SAT1 real-time RT-PCR had the highest (20%) failure rate. Briefly, topotype-specific real-time RT-PCRs had the highest serotyping capacity for EA FMDVs, although four assays were required, while the Ag-ELISA, which was less sensitive, was the most user-friendly, hence suitable for any laboratory level. In conclusion, when the four compared tests were used in combination, both the diagnostic and serotyping performances approached 100%.