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Viral Quantification in Bee Samples Using Synthetic DNA Sequences with Real-Time PCR (qPCR)

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Virus-Host Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2610))

Abstract

Pathogen spillover between honey bees and wild pollinators is a relatively new and exciting field of study. It is known that some viral diseases are a major threat to honey bee health and, thus, the diagnosis and quantification of honey bee viruses in wild pollinators have gained attention. Pathogen spillover from honey bees to wild bees and the consequences of viral replication to their health still need to be investigated. However, finding positive samples to produce standard curves and include positive controls in real-time PCR (qPCR) assays is challenging. Here we describe the use of synthetic DNA sequences of two variants of deformed wing virus (DWV-A and DWV-B), black queen cell virus (BQCV), sacbrood virus (SBV), chronic bee paralysis virus (CBPV), Kashmir bee virus (KBV), acute bee paralysis virus (ABPV), and Israeli acute paralysis virus (IAPV), to construct standard curves for viral quantification, and for their use as positive controls in qPCR assays.

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Acknowledgments

We thank Paul Kelly, Nancy Bradbury, and Stephanie Otto for their assistance during sample collection. This study was partially funded by a Pinchin Fam. Grant (Grant No. 2016030).

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Correspondence to Ernesto Guzman-Novoa .

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Morfin, N., Macías-Macías, J.O., Guzman-Novoa, E. (2023). Viral Quantification in Bee Samples Using Synthetic DNA Sequences with Real-Time PCR (qPCR). In: Aquino de Muro, M. (eds) Virus-Host Interactions. Methods in Molecular Biology, vol 2610. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2895-9_5

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  • DOI: https://doi.org/10.1007/978-1-0716-2895-9_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2894-2

  • Online ISBN: 978-1-0716-2895-9

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