QPCR Assays with Dual-labeled Probes for Genotyping Honey Bee Variants Associated with Varroa Resistance

Overview

Journal BMC Vet Res

Publisher Biomed Central

Specialty Veterinary Medicine

Date 2021 May 1

PMID 33931072

Citations 2

Authors

David Claeys Bouuaert

Mario Van Poucke

Lina De Smet

Wim Verbeke

Dirk C de Graaf

Luc Peelman

Affiliations

Soon will be listed here.

Abstract

Background: The varroa mite is one of the main causes of honey bee mortality. An important mechanism by which honey bees increase their resistance against this mite is the expression of suppressed mite reproduction. This trait describes the physiological inability of mites to produce viable offspring and was found associated with eight genomic variants in previous research.

Results: This paper presents the development and validation of high-throughput qPCR assays with dual-labeled probes for discriminating these eight single-nucleotide variants. Amplicon sequences used for assay validation revealed additional variants in the primer/probe binding sites in four out of the eight assays. As for two of these the additional variants interfered with the genotyping outcome supplementary primers and/or probes were developed. Inclusion of these primers and probes in the assay mixes allowed for the correct genotyping of all eight variants of interest within our bee population.

Conclusion: These outcomes underline the importance of checking for interfering variants in designing qPCR assays. Ultimately, the availability of this assay allows genotyping for the suppressed mite reproduction trait and paves the way for marker assisted selection in breeding programs.

Citing Articles

Allele Frequencies of Genetic Variants Associated with Varroa Drone Brood Resistance (DBR) in Subspecies across the European Continent.

Lefebre R, Smet L, Tehel A, Paxton R, Bossuyt E, Verbeke W Insects. 2024; 15(6).

PMID: 38921134 PMC: 11203681. DOI: 10.3390/insects15060419.

Population-wide modelling reveals prospects of marker-assisted selection for parasitic mite resistance in honey bees.

Lefebre R, Broeckx B, Smet L, Peelman L, de Graaf D Sci Rep. 2024; 14(1):7866.

PMID: 38570723 PMC: 10991324. DOI: 10.1038/s41598-024-58596-5.

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