非洲猪瘟病毒p30蛋白单克隆抗体的制备及阻断ELISA抗体检测方法的建立

doi:10.11843/j.issn.0366-6964.2022.12.019

Abstract

Abstract: In order to establish a blocking ELISA method for detection of African swine fever virus (ASFV) antibody, the prokaryotic expressed ASFV p30 protein was used to immunize BALB/c mice for preparation of monoclonal antibody (MAb). A blocking ELISA method was developed after optimization of the reaction conditions with the recombinant p30 protein as a coating antigen and the horseradish peroxidase (HRP) conjugated MAb as detecting antibody. The receiver operating characteristic (ROC) curve calculated an optimal cut-off value of 16.63%. This method had no cross-reaction with positive serum samples of CSFV, FMDV-O/A, PRRSV, PEDV or SVA. The sensitivity test showed that a 1∶128 dilution of positive serum can be detected by this method. The coefficient of variation (CV) of the intra- and inter-assay was less than 10%. A total of 208 serum samples were detected in parallel by the method and the commercial ELISA kit, the Kappa value was 0.96, which demonstrated its high consistency. In conclusion, the established blocking ELISA method with high sensitivity and specificity can be applied to the detection of ASFV antibodies in serum, which provides technical support for the epidemiological investigation and epidemic monitoring of ASFV.

Key words: African swine fever virus, p30 protein, monoclonal antibody, blocking ELISA

CLC Number:

S852.659.1

ZHOU Gaijing, LUO Juncong, SHI Zhengwang, WAN Ying, YANG Bo, CAO Liyan, SONG Rui, TIAN Hong, ZHENG Haixue. Preparation of Monoclonal Antibody against African Swine Fever Virus p30 Protein and Establishment of a Blocking ELISA for Detection of Antibody against ASFV p30 Protein[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4337-4345.

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Preparation of Monoclonal Antibody against African Swine Fever Virus p30 Protein and Establishment of a Blocking ELISA for Detection of Antibody against ASFV p30 Protein

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