猪伪狂犬病病毒gB蛋白抗体竞争化学发光酶联免疫检测方法的建立

doi:10.11843/j.issn.0366-6964.2020.03.017

Abstract

Abstract: A competitive and quantitative chemiluminescent enzyme immunoassay (CLEIA) for rapidly detecting antibody against gB protein of Pseudorabies Virus(PRV) was established by using the purified recombinant gB protein expressed in E. coli as coating antigen and horseradish peroxidase labelled monoclonal antibody (MAb) against gB protein as enzyme labelled antibody, and calibrator diluted from national reference to draw standard curves for quantitative detection. The successfully established PRV-gB-CLEIA which could complete the test within 45 min had no cross-reaction with the standard positive serum of other five viral antigens such as swine fever and could detect the national reference with the maximum dilution ratio of 1:2 048. The coefficient variation of intra-assay was between 1.13% and 9.47%, and inter-assay between 2.43% and 14.07%. By comparing the detection results of 180 clinical serum samples collected, the positive coincidence rate between the method and neutralization test was 94.00%, and the negative coincidence rate was 96.92%, and the total coincidence rate was 96.11%, which was obviously superior to commercial ELISA kit. The PRV-gB-CLEIA established in this study could be used for the rapid and quantitative detection of PRV gB antibody.

Key words: pseudorabies virus, gB protein, competitive chemiluminescent enzyme immunoassay, quantify

CLC Number:

S852.659.1

MA Zhenyuan, WANG Shujuan, YAN Ruoqian, BAN Fuguo, ZHAO Xueli, XIE Caihua, WANG Huajun, WANG Dongfang. Establishment of Competitive Chemiluminescent Enzyme Immunoassay for Detecting Antibodies against gB Protein of Pseudorabies Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(3): 574-583.

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Establishment of Competitive Chemiluminescent Enzyme Immunoassay for Detecting Antibodies against gB Protein of Pseudorabies Virus

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