检测结核分枝杆菌蛋白抗原b夹心ELISA的建立及其应用

发布时间：2018-06-05 15:33

本文选题：结核分枝杆菌蛋白抗原b + 结核性脑膜炎　；参考：《细胞与分子免疫学杂志》2013年05期

【摘要】：目的建立定量检测脑脊液中结核分枝杆菌蛋白抗原b的夹心ELISA,以便用于诊断结核性脑膜炎。方法制备抗结核分枝杆菌蛋白抗原b鼠源性单克隆抗体(mAb)及兔多克隆抗体;以单克隆抗体为包被抗体,多克隆抗体为检测抗体,建立检测结核分枝杆菌蛋白抗原b的双抗体夹心ELISA;应用此方法检测正常人(n=6)、非结核性脑膜炎患者(n=26)及临床确诊结核性脑膜炎患者(n=42)的脑脊液标本中蛋白抗原b的含量,并分析其与结核性脑膜炎活动性感染的相关性。结果成功建立了检测蛋白抗原b的夹心ELISA,敏感度可达0.4μg/L。应用该方法在正常人及非结核性脑膜炎患者脑脊液中未检测到蛋白抗原b;在结核性脑膜炎患者脑脊液中蛋白抗原b含量显著升高。结论建立了敏感、特异检测结核分枝杆菌蛋白抗原b含量的ELISA,为活动性结核性脑膜炎的诊断提供一种有效手段。
[Abstract]:Objective to establish a sandwich ELISA assay for the detection of Mycobacterium tuberculosis protein antigen b in cerebrospinal fluid (CSF) for the diagnosis of tuberculous meningitis. Methods the monoclonal antibody (mAb) and rabbit polyclonal antibody against Mycobacterium tuberculosis protein antigen b mouse were prepared, and the monoclonal antibody was used as the coating antibody and the polyclonal antibody as the detection antibody. To establish a double antibody sandwich ELISAmethod for detecting mycobacterium tuberculosis protein antigen b, and to detect the content of protein antigen b in cerebrospinal fluid (CSF) samples of normal persons, non-tuberculous meningitis patients, and clinically diagnosed tuberculous meningitis patients. The correlation between tuberculous meningitis and active infection was analyzed. Results the sandwich ELISAs for the detection of protein antigen b were successfully established, with a sensitivity of 0.4 渭 g / L. No protein antigen b was detected in cerebrospinal fluid (CSF) of normal subjects and non-tuberculous meningitis patients, and the content of protein antigen b in cerebrospinal fluid (CSF) of tuberculous meningitis patients increased significantly. Conclusion ELISA, which is sensitive and specific for the detection of Mycobacterium tuberculosis protein b, is an effective method for the diagnosis of active tuberculous meningitis.
【作者单位】：第四军医大学西京医院神经内科;中铁二十局中心医院;第四军医大学免疫学教研室;
【基金】：陕西省科技统筹创新工程计划(2011KTCG03-12)
【分类号】：R392.1

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