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Open AccessArticle

ABC Assay: Method Development and Application to Quantify the Role of Three DWV Master Variants in Overwinter Colony Losses of European Honey Bees

1
School of Environment and Life Sciences, The University of Salford, Manchester M5 4WT, UK
2
Viral Ecology, Marine Biological Association, Plymouth PL1 2PB, UK
3
Department of Earth, Ocean and Atmospheric Sciences and Biodiversity Research Centre, The University of British Columbia, Vancouver, BC V6T 1Z4, Canada
4
School of Biological Sciences, University of Reading, Reading RG6 6LA, UK
5
Veterinary Population Medicine, College of Veterinary Medicine, University of Minnesota, St Paul, MN 55108, USA
*
Author to whom correspondence should be addressed.
Viruses 2017, 9(11), 314; https://doi.org/10.3390/v9110314
Received: 20 September 2017 / Revised: 24 October 2017 / Accepted: 25 October 2017 / Published: 27 October 2017
(This article belongs to the Special Issue Antiviral Defense in Invertebrates)
Deformed wing virus (DWV) is one of the most prevalent honey bee viral pathogens in the world. Typical of many RNA viruses, DWV is a quasi-species, which is comprised of a large number of different variants, currently consisting of three master variants: Type A, B, and C. Little is known about the impact of each variant or combinations of variants upon the biology of individual hosts. Therefore, we have developed a new set of master variant-specific DWV primers and a set of standards that allow for the quantification of each of the master variants. Competitive reverse transcriptase polymerase chain reaction (RT-PCR) experimental design confirms that each new DWV primer set is specific to the retrospective master variant. The sensitivity of the ABC assay is dependent on whether DNA or RNA is used as the template and whether other master variants are present in the sample. Comparison of the overall proportions of each master variant within a sample of known diversity, as confirmed by next-generation sequence (NGS) data, validates the efficiency of the ABC assay. The ABC assay was used on archived material from a Devon overwintering colony loss (OCL) 2006–2007 study; further implicating DWV type A and, for the first time, possibly C in the untimely collapse of honey bee colonies. Moreover, in this study DWV type B was not associated with OCL. The use of the ABC assay will allow researchers to quickly and cost effectively pre-screen for the presence of DWV master variants in honey bees. View Full-Text
Keywords: deformed wing virus; quasi-species; honey bees; RT-qPCR; overwintering colony loss; superinfection exclusion deformed wing virus; quasi-species; honey bees; RT-qPCR; overwintering colony loss; superinfection exclusion
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Kevill, J.L.; Highfield, A.; Mordecai, G.J.; Martin, S.J.; Schroeder, D.C. ABC Assay: Method Development and Application to Quantify the Role of Three DWV Master Variants in Overwinter Colony Losses of European Honey Bees. Viruses 2017, 9, 314.

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