A new rapid and sensitive method for detecting chicken infectious anemia virus

Area of Science:

• Veterinary Virology
• Molecular Diagnostics
• Poultry Health

Background:

• Chicken infectious anemia virus (CIAV) is a globally significant immunosuppressive poultry pathogen.
• Existing detection methods for CIAV may lack the speed and accessibility required for effective on-site disease management.
• There is a critical need for novel, rapid diagnostic tools to control CIAV outbreaks in the poultry industry.

Purpose of the Study:

• To develop and validate a novel, rapid, on-site molecular diagnostic assay for chicken infectious anemia virus (CIAV).
• To establish a sensitive, specific, and cost-effective method for CIAV detection suitable for field application.
• To provide a tool for improved prevention and control strategies against CIAV in poultry.

Main Methods:

• Development of a real-time recombinase-aided amplification (RAA) assay targeting the CIAV VP2 gene.
• Optimization of reaction conditions for rapid detection at 41°C within 30 minutes.
• Validation of the assay's sensitivity, specificity, and detection limit against real-time quantitative polymerase chain reaction (qPCR).

Main Results:

• The real-time RAA assay successfully detected CIAV with a specific 117-bp band.
• The assay demonstrated a low detection limit of 10 copies of CIAV DNA per reaction.
• Comparative analysis showed high agreement with real-time qPCR (kappa=0.947), with 93.33% sensitivity and 100% specificity.

Conclusions:

• The developed real-time RAA assay is a rapid, sensitive, specific, and cost-effective method for on-site CIAV detection.
• This molecular diagnostic tool holds significant potential for routine use in CIAV prevention and control programs.
• Implementation of this assay can contribute to mitigating economic losses in the global poultry sector due to CIAV infections.