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Development and Application of an RPA-Based Rapid Point-of-Care Testing (POCT) Method for the Detection of Feline
Liang Hong1, Qian Huang1, Yuhang Zhou1
1International Immunology Centre College of Animal Science and Technology Anhui Agricultural University, Hefei 230036, Anhui, China.
Transboundary and Emerging Diseases
|April 30, 2025
Summary
A new recombinase polymerase amplification-lateral flow dipstick assay (RPA-LFDA) offers a highly sensitive and accurate method for detecting feline panleukopenia virus (FPV). This cost-effective assay is superior to existing methods for clinical diagnosis of FPV.
Area of Science:
- Veterinary Virology
- Molecular Diagnostics
- Infectious Disease Detection
Background:
- Feline panleukopenia (FP) is a severe, widespread viral disease in cats.
- Accurate and efficient detection of the feline panleukopenia virus (FPV) is crucial for timely diagnosis and treatment.
- Existing diagnostic methods may lack the desired sensitivity, specificity, or cost-effectiveness.
Purpose of the Study:
- To develop and validate a novel, rapid, and sensitive diagnostic assay for FPV detection.
- To optimize the parameters of the recombinase polymerase amplification-lateral flow dipstick assay (RPA-LFDA) for FPV.
- To evaluate the performance of the RPA-LFDA compared to established diagnostic techniques.
Main Methods:
- Development of a recombinase polymerase amplification-lateral flow dipstick assay (RPA-LFDA) for FPV detection.
- Optimization of reaction temperature (39°C) and primer selection (F3R2).
- Analytical sensitivity testing, cross-reactivity assessments with FHV-1 and FCV, and clinical validation against colloidal gold method (CGM) and qPCR.
Main Results:
- The RPA-LFDA achieved analytical sensitivity down to 10 target copies per reaction.
- No cross-reactivity was observed with feline herpesvirus-1 (FHV-1) or feline calicivirus (FCV).
- In clinical samples, RPA-LFDA demonstrated 100% sensitivity and 97.73% overall accuracy, outperforming CGM and showing high agreement with qPCR (κ=0.8962).
Conclusions:
- The developed RPA-LFDA is a highly sensitive and specific method for FPV detection.
- This assay offers advantages in convenience and cost-effectiveness over qPCR.
- RPA-LFDA presents a viable and improved option for the clinical diagnosis of feline panleukopenia virus.
