Determination of infectious bursal disease virus titration and neutralization endpoints using fluorogenic staining

Area of Science:

• Veterinary Virology
• Cell Biology
• Biotechnology

Background:

• Infectious bursal disease virus (IBDV) is a significant pathogen in poultry.
• Accurate titration and neutralization assays are crucial for IBDV diagnosis and vaccine efficacy assessment.
• Current methods can be labor-intensive and subjective.

Purpose of the Study:

• To develop and validate an automated, objective method for IBDV titration and neutralization endpoint determination.
• To compare the performance of the automated method with traditional assays.
• To establish a high-throughput screening approach for IBDV detection.

Main Methods:

• Utilized carboxyfluorescein-diacetate (CFDA) to stain cell monolayers in 96-well plates.
• Employed a fluorescence-concentration analyzer for automated data acquisition.
• Compared titration results with immunofluorescence and plaque assays.
• Determined virus-neutralization endpoints objectively.

Main Results:

• The automated method provided objective and reproducible titration and neutralization endpoints.
• Results from the automated method were comparable to immunofluorescence and plaque assays.
• Replicate tests showed equivalent or within one-dilution variability.
• Automated screening based on positive/negative controls was feasible.

Conclusions:

• The described automated method offers a reliable and objective approach for IBDV titration and neutralization assays.
• This method has the potential to improve efficiency and reduce subjectivity in IBDV diagnostics.
• The automated system facilitates high-throughput screening for IBDV detection and characterization.