Determination of parvovirus antigen in the vaccine using time-resolved fluorescence immunoassay.

Abstract

As a highly contagious and potentially fatal disease of dogs, canine parvovirus type 2 (CPV-2) usually causes severe myocarditis and gastroenteritis, while vaccine injection has greatly reduced the incidence of CPV-2 diseases. However, there is currently a lack of simple and effective method for quantitative detection of CPV-2 in vaccine. Therefore, this study aims to prepare an accurate method to determine the CPV-2 antigen (CPV-2-Ag) in vaccine. Here, a sandwich time-resolved fluorescence immunoassay (TRFIA) was established and optimized. Anti-CPV-2 antibodies were immobilized on 96-well plates to capture CPV-2-Ag, and then bound together with the detection antibodies labeled with Europium(III) (Eu3+ ) chelates; finally, time-resolved fluorometry was employed to measure the fluorescence intensity. Vaccination was performed to evaluate the relationship between CPV-2-Ag concentration and antibody titer. The sensitivity is 1.15 mEU/mL (LogY = 1.524 + 0.8667 × LogX, R2  = 0.9933), and the average recovery is among 91.00% to 106.39% without cross-reactions with the other canine viral antigen. The correlation between ELISA assay and this method is up to 0.9861. And, there is high correlation between the CPV-2-Ag concentration and antibody titers (R2  = 0.9234). This immunoassay established has high sensitivity, accuracy, and specificity, which indicate that this method could be suitable for quantitative detection of CPV-2-Ag in vaccine evaluation.

• Authors: Cuicui Chen, Huankun Liang, Baoyu Hu, Bo Ning, Hongrui Lai, Ying He, Guiling Guo, Shuhai Zhong, Laiqing Li
• Journal: Biotechnology and applied biochemistry (Biotechnol Appl Biochem) Vol. 68 Issue 3 Pg. 597-602 (Jun 2021) ISSN: 1470-8744 [Electronic] United States
• PMID: 32533780 (Publication Type: Journal Article)
• Copyright: © 2020 International Union of Biochemistry and Molecular Biology, Inc.

Chemical References

• Antibodies, Viral
• Antigens, Viral
• Viral Vaccines

Topics

• Animals
• Antibodies, Viral (immunology)
• Antigens, Viral (analysis, immunology)
• Dogs
• Enzyme-Linked Immunosorbent Assay
• Fluoroimmunoassay
• Parvovirus, Canine (immunology, isolation & purification)
• Vaccination
• Viral Vaccines (immunology)