Multiplexed detection of pathogen DNA with DNA-based fluorescence nanobarcodes.
Multiplexed detection of pathogen DNA with DNA-based fluorescence nanobarcodes.
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DOI：
10.1038/nbt1106
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发表时间：
2005-07
影响因子：
46.9
通讯作者：
Luo D
中科院分区：
文献类型：
--
作者：
Li Y;Cu YT;Luo D
在基因分析、药物筛选、临床诊断和环境分析中，对快速、多重、灵敏且特异的分子检测有很大需求。多重分析的主要挑战之一是用独特的标记或“编码”识别每个特定反应。目前使用两种编码策略：位置编码，即每种潜在反应在固相支持物（如DNA微阵列）上预先被指定一个特定位置；反应编码，即每种可能的反应都用一种通常基于光学或粒子的编码进行唯一标记。当前编码的微米尺寸、多分散性、复杂的制备过程和非生物相容性限制了它们的可用性。在此我们展示了树枝状聚合物样的基于DNA的、荧光强度编码的纳米条形码的合成，其包含内置编码和用于分子识别的探针。首先使用荧光显微镜和斑点印迹法展示了它们在多种病原体DNA的多重检测中的应用，并进一步使用流式细胞术进行了验证，结果显示检测灵敏（阿摩尔级）且快速。
本文的网络版（doi:10.1038/nbt1106）包含补充材料，授权用户可获取。
Rapid, multiplexed, sensitive and specific molecular detection is of great demand in gene profiling, drug screening, clinical diagnostics and environmental analysis. One of the major challenges in multiplexed analysis is to identify each specific reaction with a distinct label or 'code'. Two encoding strategies are currently used: positional encoding, in which every potential reaction is preassigned a particular position on a solid-phase support such as a DNA microarray, and reaction encoding, where every possible reaction is uniquely tagged with a code that is most often optical or particle based. The micrometer size, polydispersity, complex fabrication process and nonbiocompatibility of current codes limit their usability. Here we demonstrate the synthesis of dendrimer-like DNA-based, fluorescence-intensity-coded nanobarcodes, which contain a built-in code and a probe for molecular recognition. Their application to multiplexed detection of the DNA of several pathogens is first shown using fluorescence microscopy and dot blotting, and further demonstrated using flow cytometry that resulted in detection that was sensitive (attomole) and rapid. The online version of this article (doi:10.1038/nbt1106) contains supplementary material, which is available to authorized users.
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