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Updated: Aug 5, 2026
Viral Concentration Determination Through Plaque Assays: Using Traditional and Novel Overlay Systems
Published on: November 4, 2014
Quantitation of cytopathic honey bee viruses using the viral plaque assay method
Alexander McMenamin1, Michael Goblirsch2
1United States Department of Agriculture, Agricultural Research Service, Honey Bee Breeding, Genetics, and Physiology Research, Baton Rouge, LA, 78020, USA.
Abstract:
Viral pathogens are highly prevalent threats to our pollination workforce. Basic virological techniques for ascertaining infection intensity and quantifying infectious viral particles lag behind the research interest in pollinator-associated viruses. Here, we utilized the only available immortalized honey bee (Apis mellifera) cell line, AmE-711, to establish the first plaque assay protocol for a bee-infecting virus in honey bee cells. First, we demonstrate that AmE-711 is susceptible to acute bee paralysis virus (ABPV) despite being persistently infected with deformed wing virus. Then, we compared two traditional overlays, agarose and carboxymethyl cellulose (CMC) and found that the CMC method was superior in producing countable plaques in ABPV-infected monolayers. Lastly, we demonstrated that this method was useful in quantifying infectious particle loads in lysates derived from whole honey bee pupae infected with ABPV, a field-relevant sample type.
Insights
Researchers developed a new method to count infectious viral particles in honey bees. This technique uses an immortalized bee cell line and a plaque assay to better understand bee viruses and protect pollinators.
Area of Science:
- * Virology
- * Apiculture
- * Molecular Biology
Background:
- * Viral pathogens pose significant threats to honey bee (Apis mellifera) populations, impacting pollination services.
- * Current methods for quantifying bee-infecting viruses are insufficient for the growing research interest.
- * An established immortalized honey bee cell line is crucial for developing new virological assays.
Purpose of the Study:
- * To establish the first plaque assay protocol for quantifying bee-infecting viruses in an immortalized honey bee cell line.
- * To assess the susceptibility of the AmE-711 cell line to acute bee paralysis virus (ABPV).
- * To optimize the plaque assay by comparing different overlay methods and validating its use with field-relevant samples.
Main Methods:
- * Utilized the immortalized honey bee cell line AmE-711.
- * Inoculated AmE-711 cells with acute bee paralysis virus (ABPV).
- * Compared agarose and carboxymethyl cellulose (CMC) overlays for plaque formation and quantified viral loads in honey bee pupae lysates.
Main Results:
- * The AmE-711 cell line is susceptible to ABPV, even when persistently infected with deformed wing virus.
- * Carboxymethyl cellulose (CMC) overlay resulted in superior, countable plaques compared to agarose.
- * The developed plaque assay effectively quantified infectious viral particles in ABPV-infected honey bee pupae.
Conclusions:
- * A novel plaque assay protocol using the AmE-711 cell line has been established for quantifying bee-infecting viruses.
- * This assay provides a valuable tool for virological studies in honey bees, aiding in the assessment of infection intensity.
- * The method's utility in quantifying viral loads from field-relevant samples like pupae lysates will advance research on pollinator health and disease management.
