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Visual recombinase aided amplification technology for detecting feline coronavirus
Xiaoman Lu1, Yilin Cao1, Ping Zhang1
1Guangdong Laboratory for Lingnan Modern Agriculture, State Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
A new visual detection method using Recombinase Aided Amplification (RAA) rapidly and accurately identifies Feline Coronavirus (FCoV) in cats. This sensitive and specific test offers a faster, more accessible alternative for diagnosing FCoV infections.
Area of Science:
- Veterinary Diagnostics
- Molecular Biology
- Infectious Diseases
Background:
- Feline Coronavirus (FCoV) causes significant morbidity and mortality in cats globally.
- Current diagnostic methods like RT-PCR are accurate but complex and expensive.
- The lack of an effective vaccine highlights the critical need for accessible early diagnostic tools.
Purpose of the Study:
- To develop a rapid, visual, and accurate detection method for Feline Coronavirus (FCoV).
- To differentiate between FCoV serotype I and serotype II using the developed method.
- To assess the sensitivity, specificity, and clinical applicability of the novel diagnostic approach.
Main Methods:
- Utilized Recombinase Aided Amplification (RAA) technology combined with a nucleic acid-colloidal gold immunochromatographic test strip.
- Designed primers and probes targeting conserved sequences in the FCoV N and S1 genes.
- Validated the method under isothermal conditions (39°C) with a 30-minute turnaround time.
Main Results:
- The RAA method accurately identified FCoV and differentiated serotypes I and II.
- Demonstrated high specificity with no cross-reactivity against common feline viruses (FCV, FPV, FHV-1).
- Achieved superior sensitivity compared to qRT-PCR, with detection limits as low as 1.46 copies/μL for FCoV S1 gene serotype II.
Conclusions:
- The developed RAA-based visual detection method is highly accurate, sensitive, and specific for FCoV.
- Offers a rapid (30 minutes), simple, and convenient diagnostic tool for clinical settings.
- Demonstrated 100% consistency with RT-PCR in preliminary clinical applications, providing a valuable alternative for FCoV diagnosis.
