Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA, United States.
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA, United States; Emory-UGA Center of Excellence for Influenza Research and Surveillance (CEIRS), United States.
J Virol Methods. 2020 Jun;280:113878. doi: 10.1016/j.jviromet.2020.113878. Epub 2020 Apr 28.
Reassortment of segmented viruses can be an important source of genetic diversity underlying viral evolution and emergence. Methods for the quantification of reassortment have been described but are often cumbersome and best suited for the analysis of reassortment between highly divergent parental strains. While it is useful to understand the potential of divergent parents to reassort, outcomes of such heterologous reassortment are driven by differential selection acting on the progeny and are typically strain specific. To quantify reassortment robustly, a system free of differential selection is needed. We have generated such a system for influenza A virus and for mammalian orthoreovirus by constructing well-matched parental viruses carrying small genetic tags. The method utilizes high-resolution melt technology for the identification of reassortant viruses. Ease of sample preparation and data analysis enables streamlined genotyping of a large number of virus clones. The method described here thereby allows quantification of the efficiency of reassortment and can be applied to diverse segmented viruses.
重配是病毒遗传多样性的重要来源，也是病毒进化和出现的基础。目前已经有一些方法可以用于重配的定量分析，但这些方法往往比较繁琐，而且最适合用于高度分化的亲本病毒之间的重配分析。虽然了解具有较大差异的亲本病毒之间发生重配的可能性是有用的，但这种异源重配的结果是由对后代产生差异选择的作用驱动的，而且通常是针对特定的病毒株。为了稳健地定量重配，需要一个没有差异选择的系统。我们通过构建携带小遗传标记的匹配良好的亲本病毒，为甲型流感病毒和哺乳动物正呼肠孤病毒生成了这样一个系统。该方法利用高分辨率熔解技术来鉴定重组病毒。该方法易于进行样本制备和数据分析，可简化大量病毒克隆的基因分型。因此，这里描述的方法可以定量评估重配的效率，并且可以应用于不同的分段病毒。
