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目的:提取纯化结核分枝杆菌(MTB)脂阿拉伯甘露聚糖(LAM)。方法:MTB菌体彻底破碎后,去脂,去蛋白,上清液经苯酚萃取,酒精沉淀,得到LAM;以提取的LAM作为包被抗原检测血清中的LAM抗体。结果和结论:提取到LAM抗原,免疫印迹表明,LAM迁移范围相对分子质量为25×103~40×103,主要集中在35×103处。在64例肺结核患者中,有43例LAM-ELISA检测阳性(敏感性为67.19%);在67例健康志愿者中,有64例LAM-ELISA检测阴性(特异性为95.52%)。
Objective: To purify Mycobacterium tuberculosis (MTB) lipoarabinomannan (LAM). Methods: MTB cells were completely broken, defatted and deproteinized. The supernatant was extracted with phenol and precipitated with alcohol to obtain LAM. The LAM antibody was detected by using extracted LAM as coating antigen. RESULTS AND CONCLUSION: The LAM antigen was extracted and immunoblotting showed that the relative molecular mass of LAM migrated in the range of 25 × 103 ~ 40 × 103 and mainly concentrated at 35 × 103. In 64 cases of pulmonary tuberculosis, 43 cases were positive for LAM-ELISA (sensitivity was 67.19%); in 67 healthy volunteers, 64 cases were negative for LAM-ELISA (specificity was 95.52%).