Abstract：
Molecular methods are routinely used for the differential diagnosis and genetic characterization of viral disease of livestock. Real-time, quantitative PCR (qPCR) allows RNA/DNA sequence detection and quantification and is considered the gold standard diagnostic method for most viruses. However, Sanger sequencing offers additional information and opportunity to differentiate closely related virus strains and/or serotypes, by providing the full sequence of a genetic region of interest. Therefore, to determine epizootic hemorrhagic disease virus (EHDV) serotype or identify additional genetic markers, end-point RT-PCR can be performed on EHDV-positive clinical samples, followed by Sanger sequencing and data analysis. Here we describe a detailed method for the molecular characterization of EHDV serotype using Sanger sequencing.
摘要：
分子方法通常用于家畜病毒性疾病的鉴别诊断和遗传表征。实时,定量PCR（qPCR）允许RNA/DNA序列检测和定量，被认为是大多数病毒的黄金标准诊断方法。然而,Sanger测序提供了额外的信息和机会，以区分密切相关的病毒株和/或血清型，通过提供感兴趣的遗传区域的完整序列。因此,确定流行病出血性疾病病毒（EHDV）血清型或鉴定其他遗传标记，可以对EHDV阳性临床样本进行终点RT-PCR，其次是Sanger测序和数据分析。在这里，我们描述了使用Sanger测序的EHDV血清型分子表征的详细方法。
