牛传染性胸膜肺炎抗体间接ELISA检测方法的建立

doi:10.11843/j.issn.0366-6964.2020.02.014

Abstract

Abstract: An indirect ELISA was developed for detecting antibodies to Mycoplasma mycoides subsp. mycoides SC (MmmSC), the agent of contagious bovine pleuropneumonia (CBPP), and used for CBPP surveillance. Based on bioinformatics assay of the whole genome sequence of MmmSC isolates Ben-1, lipoprotein rP0308 was selected as coating antigen. An indirect ELISA was established after optimization of a series of conditions, and its performance was evaluated. As a result, the diagnostic sensitivity and specificity of the indirect ELISA were 92% and 96%, respectively. The ELISA did not react with other antibodies against Mycoplasma bovis, Mycoplasma bovirhinis, Mycoplasma agalactiae, foot and mouth disease, bovine infectious rhinotracheitis, Mycobacterium tuberculosis. Reproducibility analysis revealed that the coefficients of variation of samples within and between runs were 2.41%-6.03%, and 2.94%-6.59%, respectively. One thousand six hundred and forty-eight serum samples from our lab were tested using this ELISA system and a commercial kit, a total of 1 405/1 576 (89.1%) of the sera found positive in the commercial kit were also positive in the ELISA, and 57/72 (79.2%) of the sera found negative in the both system. The overall coincidence was 88.7%. The above results suggested that the established ELISA demonstrated high specificity, sensitivity and reproducibility, and could be a promising tool applied in the field.

Key words: contagious bovine pleuropneumonia, rP0308, indirect ELISA

CLC Number:

S852.62

HAO Wenjun, LI Yuan, ZHOU Xiaoqian, WANG Xiumei, XIN Jiuqing. Establishment of Indirect ELISA for Detecting Antibody to Contagious Bovine Pleuropneumonia[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(2): 329-336.

References 20

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Establishment of Indirect ELISA for Detecting Antibody to Contagious Bovine Pleuropneumonia

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