脂阿拉伯甘露聚糖的提纯及其对肺结核的诊断价值

目的 初步建立提取纯化结核分枝杆菌（M .TB）脂阿拉伯甘露聚糖（lipoarabinomanan,LAM）的方法 ，并将其用于血清标本检测，评价其对肺结核的诊断价值。 方法 M .TB菌体经冰浴超声破碎，乙醇回流，DNase I 酶RNase酶除去DNA和RNA, 热酚水法去蛋白，氯仿甲醇去酚和脂质，得到粗制的脂多糖。经Sephacryl-100凝胶过滤层析分离纯化,得到LAM。将其作为抗原，通过免疫金渗滤法检测涂阳肺结核和对照血清标本中的LAM抗体。 结果 所得LAM经SDS-PAGE银染，与对照标准品大小一致，为37 kDa。血清LAM抗体检测对肺结核诊断的敏感性、特异性、阳性预测值、阴性预测值、和涂片符合率分别为91.7%、87.8%、92.7%、86.2%、90.2%。 结论 成功提取到LAM抗原，用其检测血清结核抗体显示出理想的敏感性和特异性，有望成为结核病血清学快速诊断方法之一。

Extraction of lipoarabinomanan and its diagnosis value for pulmonary tuberculosis

Objective To develop a method for extracting lipoarabinomanan(LAM) from Mycobacterium tuberculosis,and to evaluate serodiagnosis value of LAM for pulmonary tuberculosis.Methods Mycobacterium tuberculosis was sonicated in ice and then refluxed with ethanol;treated with DNase I and RNase;removed proteins with phenol;removed phenol and lipide with chloroform/methanol.The crude lipopolysaccharide(LPS) was prepared.LPS was purified with Sephacryl-100 gel filtration,and LAM were obtained.LAM was used as antigen t...