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Development and Validation of a Recombinant VP2-Based Indirect ELISA for Canine Parvovirus
Bocheng Gao1, Jiale Yi2, Linna Gai1
1Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, College of Bioscience and Engineering, Institute of Pathogenic Microorganism, Jiangxi Agricultural University, Nanchang 330045, China.
Microorganisms
|May 27, 2026
Summary
Researchers developed a new indirect ELISA test using recombinant canine parvovirus (CPV) VP2 protein to detect CPV-specific antibodies in dog serum. This assay demonstrates good sensitivity and specificity, offering a reliable diagnostic tool for canine parvovirus.
Area of Science:
- Veterinary Virology
- Immunodiagnostics
- Molecular Biology
Background:
- Canine parvovirus (CPV) poses a significant threat to canine health.
- Accurate detection of CPV-specific antibodies is crucial for disease management and vaccination assessment.
- Existing diagnostic methods may have limitations in sensitivity or accessibility.
Purpose of the Study:
- To express the canine parvovirus (CPV) VP2 protein prokaryotically.
- To develop and validate an indirect enzyme-linked immunosorbent assay (ELISA) for detecting CPV-specific antibodies in canine serum.
Main Methods:
- Amplification and cloning of the CPV VP2 gene.
- Prokaryotic expression and purification of recombinant VP2 protein using Ni-NTA affinity chromatography.
- Development of an indirect ELISA with optimized antigen concentration and antibody dilutions.
- Validation using Western blotting and comparison with the hemagglutination inhibition (HI) assay.
Main Results:
- Successfully expressed and purified a 64.8 kDa recombinant CP2 protein.
- Established an indirect ELISA with optimal parameters: antigen coating (2 μg/mL), serum dilution (1:320), and secondary antibody dilution (1:4000).
- The ELISA demonstrated high analytical sensitivity (1:5120 dilution), good diagnostic sensitivity (85.94%), specificity (88.28%), and overall agreement (87.50%) compared to the HI assay.
Conclusions:
- The recombinant VP2-based indirect ELISA is a sensitive, reproducible, and reliable method for detecting CPV-specific antibodies in canine serum.
- This assay shows significant diagnostic agreement with the established HI assay.
- Further validation for broader cross-reactivity is recommended.
