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Updating Sacbrood Virus Quantification PCR Method Using a TaqMan-MGB Probe.

Wei-Fone Huang1, Yakun Zhang1, Shahid Mehmood2

  • 1College of Animal Science (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, China.

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Summary

Updated qPCR methods can now detect Sacbrood virus (SBV) variants in Asian honey bees (Apis cerana). This is crucial as these variants can infect European honey bees (Apis mellifera) and other insects.

Keywords:
AcSBVCSBVMGB probeTaqManThai SBV

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Area of Science:

  • Veterinary Virology
  • Apiculture Science
  • Molecular Diagnostics

Background:

  • Sacbrood virus (SBV) is a significant pathogen affecting honey bee populations globally.
  • Distinct SBV variants in Asian honey bees (Apis cerana) pose identification challenges and may evade current detection methods.
  • The potential for cross-species transmission of SBV variants to European honey bees (Apis mellifera) necessitates accurate diagnostic tools.

Purpose of the Study:

  • To update and validate a quantitative PCR (qPCR) method for detecting Sacbrood virus (SBV) variants, including those from Asian honey bees (Apis cerana).
  • To ensure the updated method can accurately identify newly emerged SBV variants (Ac SBV) that may not be recognized by existing assays.
  • To provide a reliable diagnostic tool for monitoring SBV in various hymenopteran species.

Main Methods:

  • The study updated a TaqMan qPCR assay based on Blanchard's design, utilizing conserved regions of the VP1 gene.
  • Minor groove binder (MGB) probes were incorporated for enhanced detection specificity.
  • The updated method was validated using samples from European honey bees (Apis mellifera), Asian honey bees (Apis cerana), and the hornet species Vespa velutina.

Main Results:

  • The updated primers and probe demonstrated compatibility with published SBV and Ac SBV genome sequences in databases.
  • The revised qPCR method exhibited reasonable sensitivity and flexibility for detecting and quantifying SBV.
  • The assay successfully detected SBV in tested bee and hornet species, including Apis mellifera and Apis cerana.

Conclusions:

  • The developed qPCR method provides an improved tool for the detection and quantification of Sacbrood virus (SBV) variants, including those originating from Asian honey bees.
  • This updated assay is essential for monitoring SBV epidemiology and managing its impact on honey bee health, especially considering potential cross-species infections.
  • The method's flexibility allows for its application in detecting SBV in Apis mellifera, Apis cerana, and Vespa velutina, pending the emergence of variants with significantly mutated VP1 genes.