1.Establishment of a monoclonal antibody-based competitive chemiluminescent en-zyme-linked immunosorbent assay for detection of Senecavirus A antibodies
Zhenyuan MA ; Ruoqian YAN ; Mao CHAI ; Shujuan WANG ; Xueli ZHAO ; Haibo YANG ; Dongfang WANG ; Ying LIU ; Cui WANG
Chinese Journal of Veterinary Science 2025;45(7):1402-1410
In order to establish a competitive chemiluminescent enzyme-linked immunoassay for rapid and quantitative detection of Senecavirus A antibodies,the polystyrene plate was coated with inactivated Senecavirus A antigen,and the monoclonal antibodies against Senecavirus A VP2 and VP3 proteins labeled by horseradish peroxidase(HRP)were used as the competitive enzymic anti-bodies of the antibodies in the serum samples.The standard curve of the calibrator prepared by di-lution of positive serum was drawn to achieve quantitative detection.The successfully established SVA competitive CLEIA reported the result within 45 minutes.The maximum dilution of 1∶2 048 for calibrator serum was still detectable with no cross-reaction with the standard positive serum of other five kinds of virus antigens such as foot-and-mouth disease.The coefficient of variation within batches was less than 10%,and the coefficient of variation between batches was less than 15%,which showed good repeatability and stability.The positive and negative coincidence rates were 95.30%and 97.57%,respectively,and the total coincidence rate was 96.88%,showing high consistency.The SVA competitive CLEIA assay established in this study can be used for the rapid quantitative detection of Senecavirus A antibodies,filling the gap in the domestic rapid quantitative detection of SVA antibodies.
2.Establishment of a monoclonal antibody-based competitive chemiluminescent en-zyme-linked immunosorbent assay for detection of Senecavirus A antibodies
Zhenyuan MA ; Ruoqian YAN ; Mao CHAI ; Shujuan WANG ; Xueli ZHAO ; Haibo YANG ; Dongfang WANG ; Ying LIU ; Cui WANG
Chinese Journal of Veterinary Science 2025;45(7):1402-1410
In order to establish a competitive chemiluminescent enzyme-linked immunoassay for rapid and quantitative detection of Senecavirus A antibodies,the polystyrene plate was coated with inactivated Senecavirus A antigen,and the monoclonal antibodies against Senecavirus A VP2 and VP3 proteins labeled by horseradish peroxidase(HRP)were used as the competitive enzymic anti-bodies of the antibodies in the serum samples.The standard curve of the calibrator prepared by di-lution of positive serum was drawn to achieve quantitative detection.The successfully established SVA competitive CLEIA reported the result within 45 minutes.The maximum dilution of 1∶2 048 for calibrator serum was still detectable with no cross-reaction with the standard positive serum of other five kinds of virus antigens such as foot-and-mouth disease.The coefficient of variation within batches was less than 10%,and the coefficient of variation between batches was less than 15%,which showed good repeatability and stability.The positive and negative coincidence rates were 95.30%and 97.57%,respectively,and the total coincidence rate was 96.88%,showing high consistency.The SVA competitive CLEIA assay established in this study can be used for the rapid quantitative detection of Senecavirus A antibodies,filling the gap in the domestic rapid quantitative detection of SVA antibodies.

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