Functional Genome Analysis, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 580, 69120 Heidelberg, Germany.
J Proteome Res. 2011 Mar 4;10(3):1316-22. doi: 10.1021/pr101070j. Epub 2011 Feb 16.
Based on a single-molecule sensitive fluorescence-linked immunosorbent assay, an analytical platform for the detection of lipoarabinomannan (LAM), a lipopolysaccharide marker of tuberculosis, was established that is about 3 orders of magnitude more sensitive than comparable current ELISA assays. No amplification step was required. Also, no particular sample preparation had to be done. Since individual binding events are detected, true quantification was possible simply by counting individual signals. Utilizing a total internal reflection configuration, unprocessed biological samples (human urine and plasma) to which LAM was added could be analyzed without the requirement of sample purification or washing steps during analysis. Samples containing about 600 antigen molecules per microliter produced a distinct signal. The methodology developed can be employed for any set of target molecules for which appropriate antibodies exist.
基于单分子敏感荧光酶联免疫吸附测定法，建立了一种用于检测脂阿拉伯甘露聚糖 (LAM) 的分析平台，其灵敏度比可比的当前 ELISA 测定法高约 3 个数量级。该方法不需要扩增步骤，也不需要特殊的样品制备。由于检测到的是单个结合事件，因此只需通过计数单个信号即可进行真正的定量。利用全内反射配置，可以分析添加了 LAM 的未处理生物样品（人尿和血浆），而无需在分析过程中进行样品纯化或洗涤步骤。每个微升中含有约 600 个抗原分子的样品会产生明显的信号。所开发的方法可用于任何具有合适抗体的目标分子组。
