A method for the unbiased quantification of reassortment in segmented viruses

• Type of Material: Article
• Authors: Megan R Hockman, Emory University; Kara L Phipps, Emory University; Katie E Holmes, Emory University; Anice Lowen, Emory University
• Language: English
• Date: 2020-06-01
• Publisher: ELSEVIER
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2020 Elsevier B.V. All rights reserved.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.jviromet.2020.113878
• Title of Journal or Parent Work: JOURNAL OF VIROLOGICAL METHODS
• Volume: 280
• Start Page: 113878
• End Page: 113878
• Grant/Funding Information: This work was funded in part by the NIH/NIAID Centers of Excellence for Influenza Research and Surveillance (CEIRS) contract HHSN272201400004C and NIH grant R01AI125268 to AL.
• Keywords: Virology; Reassortment; FEATURES; INFECTION; Science & Technology; Mammalian orthoreovirus; REPLICATION; Biochemical Research Methods; Biotechnology & Applied Microbiology; Viral genetics; INFLUENZA-A VIRUS; Biochemistry & Molecular Biology; Influenza A virus; POLYPEPTIDES; Life Sciences & Biomedicine; SEGREGATION; PROTEIN; REVERSE GENETICS; TEMPERATURE-SENSITIVE MUTANTS; ORIGIN

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.