脂阿拉伯甘露聚糖的提纯及其对肺结核的诊断价值  

Extraction of lipoarabinomanan and its diagnosis value for pulmonary tuberculosis

作　　者：彭荣[1] 乐军[1] 金瑞良[1] 景玲杰[1] 韩敏[1] 

机构地区：[1]同济大学附属上海市肺科医院/上海市结核(肺)重点实验室,上海200433

出　　处：《中国防痨杂志》2011年第3期149-152,共4页Chinese Journal of Antituberculosis

基　　金：国家"十一五"科技重大专项(2008ZX10003003);上海市卫生局青年基金(2008Y26)

摘　　要：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。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 to detect anti-LAM IgG in serum by dot immunogold filtration assay.Results The relative molecular weight of LAM was approximately 37 kDa,consistent with the reference.The sensitivity,specificity,positive predictive value,negative predictive value and coincidence with smear were 91.7%,87.8%,92.7%,86.2% and 90.2%,respectively.Conclusions LAM was obtained successfully.LAM used as antigen to detect anti-TB antibody in serum showed good specificity and sensitivity,and might be one of canditate antigens for the serodiagnosis of TB serology.

关 键 词：结核 肺/诊断 分枝杆菌 结核 脂多糖类 

分 类 号：R521[医药卫生—内科学]