Intra-laboratory validation of chronic bee paralysis virus quantitation using an accredited standardised real-time...

Area of Science:

• Veterinary Virology
• Apiculture Science
• Molecular Diagnostics

Background:

• Chronic Bee Paralysis Virus (CBPV) causes a significant infectious disease in adult honey bees (Apis mellifera L.).
• Accurate quantification of CBPV is essential for understanding disease dynamics and implementing control strategies.
• Existing diagnostic methods may lack the precision required for reliable CBPV load assessment.

Purpose of the Study:

• To characterize and validate a real-time RT-PCR assay for the accurate quantification of CBPV.
• To assess the reliability and repeatability of the assay as a potential reference method.
• To ensure the assay meets international standards for diagnostic test validation.

Main Methods:

• Intra-laboratory characterization of a real-time RT-PCR assay for CBPV.
• Performance assessment of both the qPCR assay and the complete RNA extraction-to-analysis method.
• Validation according to ISO/IEC 17025 and XP U47-600 standards.

Main Results:

• The real-time RT-PCR assay demonstrated high reliability and repeatability.
• The assay and overall method were validated over a 6 log range (10^2 to 10^8 copies).
• A detection limit of 50-100 CBPV RNA copies was achieved, confirming assay sensitivity.

Conclusions:

• The characterized real-time RT-PCR assay provides a reliable method for CBPV quantitation.
• The validated protocol is suitable for use as a reference method in diagnostic laboratories.
• The assay's approval by the French Accreditation Committee underscores its robustness for CBPV monitoring.