Application of high-throughput sequencing to whole rabies viral genome characterisation and its use for phylogenetic re-evaluation of a raccoon strain incursion into the province of Ontario

• Authors: Susan A. Nadin-Davis, Canadian Food Inspection Agency; Adam Colville, Canadian Food Inspection Agency; Hannah Trewby, University of Glasgow; Roman Biek, University of Glasgow; Leslie Real, Emory University
• Journal Title: Virus Research
• Volume: Volume 232
• Publisher: Elsevier | 2017-03-15, Pages 123-133
• Type of Work: Article | Final Publisher PDF
• Publisher DOI: http://doi.org/10.1016/j.virusres.2017.02.007
• Subjects: Biology, Microbiology; Biology, General
• Keywords: raccoon rabies virus; Illumina next generation sequencing; molecular epidemiology; viral phylogeny

Abstract

Raccoon rabies remains a serious public health problem throughout much of the eastern seaboard of North America due to the urban nature of the reservoir host and the many challenges inherent in multi-jurisdictional efforts to administer co-ordinated and comprehensive wildlife rabies control programmes. Better understanding of the mechanisms of spread of rabies virus can play a significant role in guiding such control efforts. To facilitate a detailed molecular epidemiological study of raccoon rabies virus movements across eastern North America, we developed a methodology to efficiently determine whole genome sequences of hundreds of viral samples. The workflow combines the generation of a limited number of overlapping amplicons covering the complete viral genome and use of high throughput sequencing technology. The value of this approach is demonstrated through a retrospective phylogenetic analysis of an outbreak of raccoon rabies which occurred in the province of Ontario between 1999 and 2005. As demonstrated by the number of single nucleotide polymorphisms detected, whole genome sequence data were far more effective than single gene sequences in discriminating between samples and this facilitated the generation of more robust and informative phylogenies that yielded insights into the spatio-temporal pattern of viral spread. With minor modification this approach could be applied to other rabies virus variants thereby facilitating greatly improved phylogenetic inference and thus better understanding of the spread of this serious zoonotic disease. Such information will inform the most appropriate strategies for rabies control in wildlife reservoirs.