Knowles N J, Wadsworth J, Bachanek-Bankowska K, King D P
Rev Sci Tech. 2016 Dec;35(3):741-755. doi: 10.20506/rst.35.3.2565.
Nucleotide sequences of field strains of foot and mouth disease virus (FMDV) contribute to our understanding of the distribution and evolution of viral lineages that circulate in different regions of the world. This paper outlines a practical reversetranscription polymerase chain reaction (RT-PCR) and sequencing strategy that can be used to generate RNA sequences encoding the VP1 (1D) region of FMDV. The protocol contains a panel of PCR and sequencing primers that can be selected to characterise genetically diverse isolates representing all seven FMDV serotypes. A list of sequences is also described, comprising prototype sequences for all proposed FMDV topotypes, in order to provide a framework for phylogenetic analysis. The technical details and prototype sequences provided in this paper can be employed by FMD Reference Laboratories and others in an approach to harmonise the molecular epidemiology of FMDV.
口蹄疫病毒(FMDV)田间毒株的核苷酸序列有助于我们了解在世界不同地区传播的病毒谱系的分布和进化情况。本文概述了一种实用的逆转录聚合酶链反应(RT-PCR)和测序策略,可用于生成编码FMDV VP1(1D)区域的RNA序列。该方案包含一组PCR和测序引物,可用于对代表所有七种FMDV血清型的基因多样化分离株进行特征分析。还描述了一个序列列表,包括所有提议的FMDV拓扑型的原型序列,以便为系统发育分析提供框架。本文提供的技术细节和原型序列可供FMD参考实验室及其他机构采用,以统一FMDV的分子流行病学研究方法。