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qPCR assays with dual-labeled probes for genotyping honey bee variants associated with varroa resistance

David Claeys Boúúaert (UGent) , Mario Van Poucke (UGent) , Lina De Smet (UGent) , Wim Verbeke (UGent) , Dirk de Graaf (UGent) and Luc Peelman (UGent)
(2021) BMC VETERINARY RESEARCH. 17(1).
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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.
Keywords
General Veterinary, General Medicine, Honey bee, Varroa destructor, Varroa resistance, Suppressed mite reproduction, Resilience, High-throughput DNA test, DESTRUCTOR, SELECTION, MITE

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MLA
Claeys Boúúaert, David, et al. “QPCR Assays with Dual-Labeled Probes for Genotyping Honey Bee Variants Associated with Varroa Resistance.” BMC VETERINARY RESEARCH, vol. 17, no. 1, 2021, doi:10.1186/s12917-021-02886-x.
APA
Claeys Boúúaert, D., Van Poucke, M., De Smet, L., Verbeke, W., de Graaf, D., & Peelman, L. (2021). qPCR assays with dual-labeled probes for genotyping honey bee variants associated with varroa resistance. BMC VETERINARY RESEARCH, 17(1). https://doi.org/10.1186/s12917-021-02886-x
Chicago author-date
Claeys Boúúaert, David, Mario Van Poucke, Lina De Smet, Wim Verbeke, Dirk de Graaf, and Luc Peelman. 2021. “QPCR Assays with Dual-Labeled Probes for Genotyping Honey Bee Variants Associated with Varroa Resistance.” BMC VETERINARY RESEARCH 17 (1). https://doi.org/10.1186/s12917-021-02886-x.
Chicago author-date (all authors)
Claeys Boúúaert, David, Mario Van Poucke, Lina De Smet, Wim Verbeke, Dirk de Graaf, and Luc Peelman. 2021. “QPCR Assays with Dual-Labeled Probes for Genotyping Honey Bee Variants Associated with Varroa Resistance.” BMC VETERINARY RESEARCH 17 (1). doi:10.1186/s12917-021-02886-x.
Vancouver
1.
Claeys Boúúaert D, Van Poucke M, De Smet L, Verbeke W, de Graaf D, Peelman L. qPCR assays with dual-labeled probes for genotyping honey bee variants associated with varroa resistance. BMC VETERINARY RESEARCH. 2021;17(1).
IEEE
[1]
D. Claeys Boúúaert, M. Van Poucke, L. De Smet, W. Verbeke, D. de Graaf, and L. Peelman, “qPCR assays with dual-labeled probes for genotyping honey bee variants associated with varroa resistance,” BMC VETERINARY RESEARCH, vol. 17, no. 1, 2021.
@article{8737150,
  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.}},
  articleno    = {{179}},
  author       = {{Claeys Boúúaert, David and Van Poucke, Mario and De Smet, Lina and Verbeke, Wim and de Graaf, Dirk and Peelman, Luc}},
  issn         = {{1746-6148}},
  journal      = {{BMC VETERINARY RESEARCH}},
  keywords     = {{General Veterinary,General Medicine,Honey bee,Varroa destructor,Varroa resistance,Suppressed mite reproduction,Resilience,High-throughput DNA test,DESTRUCTOR,SELECTION,MITE}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{6}},
  title        = {{qPCR assays with dual-labeled probes for genotyping honey bee variants associated with varroa resistance}},
  url          = {{http://dx.doi.org/10.1186/s12917-021-02886-x}},
  volume       = {{17}},
  year         = {{2021}},
}
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