Development of a direct competitive ELISA for the detection of Mycoplasma bovis infection based on a monoclonal antibody of P48 protein.
Development of a direct competitive ELISA for the detection of Mycoplasma bovis infection based on a monoclonal antibody of P48 protein.
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DOI：
10.1186/1746-6148-10-42
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发表时间：
2014-02-18
影响因子：
2.6
通讯作者：
Wu W
中科院分区：
文献类型：
--
作者：
Fu P;Sun Z;Zhang Y;Yu Z;Zhang H;Su D;Jiang F;Wu W
牛支原体（M.牛（bovis）是一种主要但经常被忽视的病原体，自2008年以来，在中国各地记录了导致牛呼吸道疾病、乳腺炎和关节炎的病原体。本文报道了一种直接竞争酶联免疫吸附试验（Dc-ELISA）检测支原体的方法。牛抗体我们使用重组P48蛋白和单克隆抗体（mAb）10 E。单克隆抗体10 E，针对M. bovis，鉴定出所有M.与其他相关病原体无交叉反应的牛种。用P48蛋白代替全M包被微孔板。牛细胞以及使用mAb 10 E产生了特异性和敏感性的M.牛抗体检测，截止抑制百分比（PI）值为32%。与两种商品化的间接ELISA（i-ELISA）试剂盒相比，Dc-ELISA对165份临床牛血清样品的阳性检出率较高。建立了一种快速、敏感、可靠的M. bovis，可促进M.牛的监测，协助研究人员了解生态和流行病学的M。牛。
Mycoplasma bovis (M. bovis) is a major, but often overlooked, pathogen documented to cause respiratory disease, mastitis, and arthritis in cattle throughout China since 2008. Here, we report the development of a direct competitive enzyme-linked immunosorbent assay (Dc-ELISA) to detect M. bovis antibody. We used a recombinant P48 protein and monoclonal antibody (mAb) 10E. MAb 10E, prepared against the recombinant P48 protein of M. bovis, identified all M. bovis strains with no cross-reactivity with other related pathogens. Coating micro plates with P48 protein instead of whole M. bovis cells as well as the use of mAb 10E produced a specific and sensitive Dc-ELISA for M. bovis antibody detection with a cut-off percent inhibition (PI) value of 32%. Compared with two commercial indirect ELISA (i-ELISA) kits, our Dc-ELISA offered a higher positive detection rate in 165 clinical bovine serum samples. A rapid, sensitive, and reliable serological diagnosis method was developed for M. bovis, which can facilitate M. bovis surveillance, assisting researchers in understanding the ecology and epidemiology of M. bovis.
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