A method for the unbiased quantification of reassortment in segmented viruses

• Authors: Megan R Hockman, Emory University; Kara L Phipps, Emory University; Katie E Holmes, Emory University; Anice Lowen, Emory University
• Journal Title: JOURNAL OF VIROLOGICAL METHODS
• Volume: Volume 280
• Publisher: ELSEVIER | 2020-06-01, Pages 113878-113878
• Type of Work: Article | Post-print: After Peer Review
• Publisher DOI: http://doi.org/10.1016/j.jviromet.2020.113878
• Subjects: Health Sciences, Immunology
• Keywords: Science & Technology; Life Sciences & Biomedicine; Biochemical Research Methods; Biotechnology & Applied Microbiology; Virology; Biochemistry & Molecular Biology; Reassortment; Mammalian orthoreovirus; Influenza A virus; Viral genetics; INFLUENZA-A VIRUS; TEMPERATURE-SENSITIVE MUTANTS; REVERSE GENETICS; ORIGIN; POLYPEPTIDES; REPLICATION; SEGREGATION; INFECTION; FEATURES; PROTEIN

Abstract

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.