A quantitative loop-mediated isothermal amplification assay for detecting a novel goose astrovirus

Area of Science:

• Veterinary Virology
• Molecular Diagnostics
• Avian Infectious Diseases

Background:

• A novel goose astrovirus (N-AstV) caused a severe infectious disease in China in 2017.
• The disease outbreaks significantly impacted goose production, necessitating rapid diagnostic methods.

Purpose of the Study:

• To develop a quantitative loop-mediated isothermal amplification (qLAMP) assay for N-AstV detection.
• To provide a rapid, sensitive, and specific diagnostic tool for N-AstV.

Main Methods:

• Designed 4 specific primers (2 inner, 2 outer) targeting the N-AstV ORF1a gene.
• Developed a qLAMP assay for N-AstV diagnosis, executable within 60 minutes at 65°C.
• Validated assay sensitivity, specificity, and reproducibility using laboratory tests.

Main Results:

• The qLAMP assay demonstrated high sensitivity with a detection limit of 1 × 10^1 copies of target DNA/μL.
• No cross-reactions were observed with other viruses, confirming specificity.
• High reproducibility was confirmed with variability ranging from 0.61% to 2.21%.

Conclusions:

• The developed qLAMP assay is a simple, accurate, rapid, sensitive, and specific method for N-AstV detection.
• This assay is particularly valuable for field-based diagnosis and disease control in goose populations.