A semi-automated and high-throughput approach for the detection of honey bee viruses in bee samples

Author:

Nikulin Sofia Levin,Hesketh-Best Poppy J.ORCID,Mckeown Dean A.,Spivak Marla,Schroeder Declan C.ORCID

Abstract

Deformed wing virus (DWV) was first detected in dead honey bees in 1982 but has been in honey bees for at least 300 years. Due to its high prevalence and virulence, they have been linked with the ongoing decline in honey bee populations worldwide. A rapid, simple, semi-automated, high-throughput, and cost-effective method of screening colonies for viruses would benefit bee research and the beekeeping industry. Here we describe a semi-automated approach that combines an RNA-grade liquid homogenizer followed by magnetic bead capture for total virus nucleic acid extraction. We compare it to the more commonly applied nucleic acid column-based purification method and use qPCR plus Oxford Nanopore Technologies sequencing to evaluate the accuracy of analytical results for both methods. Our results showed high reproducibility and accuracy for both approaches. The semi-automated method described here allows for faster screening of viral loads in units of 96 samples at a time. We developed this method to monitor viral loads in honey bee colonies, but it could be easily applied for any PCR or genomic-based screening assays.

Funder

National Institute of Food and Agriculture

Minnesota Environment and Natural Resources Trust Fund

Publisher

Public Library of Science (PLoS)

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