Development and application of a blocking ELISA based on a N protein monoclonal antibody for the antibody detection against porcine reproductive and respiratory syndrome virus 2
文献类型： 外文期刊
作者： Li, Wansheng 1 ; Li, Yanwei 2 ; Li, Minhua 3 ; Zhang, Hongliang 1 ; Feng, Zixuan 1 ; Xu, Hu 1 ; Li, Chao 1 ; Guo, Zhenyang 1 ; Gong, Bangjun 1 ; Peng, Jinmei 1 ; Zhou, Guohui 1 ; Tian, Zhijun 1 ; Wang, Qian 1 ;
作者机构： 1.Chinese Acad Agr Sci, Harbin Vet Res Inst, State Key Lab Anim Dis Control & Prevent, Harbin 150001, Peoples R China
2.Nanjing Co Ltd, Beijing Biomed Technol Ctr JoFunHwa Biotechnol, Beijing 102600, Peoples R China
3.Beijing IDEXX Labs Co Ltd, Beijing 101318, Peoples R China
4.CAAS, Harbin Vet Res Inst, 678 Haping Rd, Harbin 150069, Peoples R China
关键词： Porcine reproductive and respiratory syndrome; virus-2; N protein monoclonal antibody; Blocking ELISA
期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.2； 五年影响因子：7.8 ）
ISSN： 0141-8130
年卷期： 2024 年 269 卷
页码：
收录情况： SCI
摘要： Porcine reproductive and respiratory syndrome (PRRS) is one of the most widespread illnesses in the world's swine business. To detect the antibodies against PRRSV-2, a blocking enzyme-linked immunosorbent assay (BELISA) was developed, utilizing a PRRSV-2 N protein monoclonal antibody as the detection antibody. A checkerboard titration test was used to determine the optimal detection antibody dilution, tested pig serum dilution and purified PRRSV coated antigen concentration. After analyzing 174 negative pig sera and 451 positive pig sera, a cutoff value of 40 % was selected to distinguish between positive and negative sera using receiver operating characteristic curve analysis. The specificity and sensitivity of the assay were evaluated to equal 99.8 % and 96 %, respectively. The method had no cross-reaction with PCV2, PRV, PPV, CSFV, PEDV, TGEV, and PRRSV-1 serum antibodies, and the coefficients of variation of intra-batch and inter-batch repeatability experiments were both <10 %. A total of 215 clinical serum samples were tested, and the relative coincidence rate with commercial ELISA kit was 99.06 %, and the kappa value was 0.989, indicating that these two detection results exhibited high consistency. Overall, the B-ELISA should serve as an ideal method for large-scale serological investigation of PRRSV-2 antibodies in domestic pigs.
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