Homogeneous detection of unamplified genomic DNA sequences based on colorimetric scatter of gold nanoparticle probes
Homogeneous detection of unamplified genomic DNA sequences based on colorimetric scatter of gold nanoparticle probes
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DOI：
10.1038/nbt977
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发表时间：
2004-07-01
影响因子：
46.9
通讯作者：
Müller, UR
中科院分区：
文献类型：
--
作者：
Storhoff, JJ;Lucas, AD;Müller, UR
核酸诊断主要是基于荧光的测定，其使用复杂且昂贵的基于酶的靶或信号放大程序（1-6）。许多临床诊断应用将需要可以在筛选模式下进行的更简单、便宜的测定。我们已经开发了一种基于金纳米颗粒的距离依赖性光学性质的“点读”比色检测方法用于识别核酸序列。在该测定中，核酸靶标被DNA修饰的金探针识别，当溶液点样到照明的玻璃波导上时，所述金探针经历可目视检测的颜色变化。这种基于散射的方法能够在与促进探针-靶标以均质形式结合的改进的杂交方法结合时，在没有靶标或信号扩增的情况下检测zeptomole量的核酸靶标。与先前报道的基于吸光度的方法7相比，该方法将检测灵敏度提高了四个数量级以上。我们已将此方法应用于耐甲氧西林金黄色葡萄球菌基因组DNA样品中mecA的快速检测。
Nucleic acid diagnostics is dominated by fluorescence-based assays that use complex and expensive enzyme-based target or signal-amplification procedures(1-6). Many clinical diagnostic applications will require simpler, inexpensive assays that can be done in a screening mode. We have developed a 'spot-and-read' colorimetric detection method for identifying nucleic acid sequences based on the distance-dependent optical properties of gold nanoparticles. In this assay, nucleic acid targets are recognized by DNA-modified gold probes, which undergo a color change that is visually detectable when the solutions are spotted onto an illuminated glass waveguide. This scatter-based method enables detection of zeptomole quantities of nucleic acid targets without target or signal amplification when coupled to an improved hybridization method that facilitates probe-target binding in a homogeneous format. In comparison to a previously reported absorbance-based method 7, this method increases detection sensitivity by over four orders of magnitude. We have applied this method to the rapid detection of mecA in methicillin-resistant Staphylococcus aureus genomic DNA samples.