Establishment of a Blocking ELISA Detection Method for Against African Swine Fever Virus p30 Antibody

• Published in: Frontiers in veterinary science 2021-12, Vol.8, p.781373-781373
• Main Authors: Yu, Xuexiang, Zhu, Xianjing, Chen, Xiaoyu, Li, Dongfan, Xu, Qian, Yao, Lun, Sun, Qi, Ghonaim, Ahmed H, Ku, Xugang, Fan, Shengxian, Yang, Hanchun, He, Qigai
• Format: Article
• Language: English
• Subjects: African swine fever virus; blocking ELISA; diagnosis; monoclonal antibodies; p30; Veterinary Science
• ISSN: 2297-1769; 2297-1769
• DOI: 10.3389/fvets.2021.781373

African swine fever (ASF) is a highly lethal hemorrhagic viral disease of domestic pigs caused by African swine fever virus (ASFV). A sensitive and reliable serological diagnostic assay is required, so laboratories can effectively and quickly detect ASFV infection. The p30 protein is abundantly expressed early in cells and has excellent antigenicity. Therefore, this study aimed to produce and characterize p30 monoclonal antibodies with an ultimate goal of developing a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) for ASFV antibody detection. Three monoclonal antibodies against p30 protein that were expressed in were generated, and their characterizations were investigated. Furthermore, a blocking ELISA based on a monoclonal antibody was developed. To evaluate the performance of the assay, 186 sera samples (88 negative and 98 positive samples) were analyzed and a receiver-operating characteristic (ROC) analysis was applied to determine the cutoff value. Based on the ROC analysis, the area under the curve (AUC) was 0.997 (95% confidence interval: 99.2 to 100%). Besides, a diagnostic sensitivity of 97.96% (95% confidence interval: 92.82 to 99.75%) and a specificity of 98.96% (95% confidence interval: 93.83 to 99.97%) were achieved when the cutoff value was set to 38.38%. Moreover, the coefficients of inter- and intra-batches were