Base pair mismatch recognition using plasmon resonant particle labels.
Base pair mismatch recognition using plasmon resonant particle labels.
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DOI：
10.1016/s0003-2697(02)00410-4
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发表时间：
2002-10
影响因子：
2.9
通讯作者：
S. Oldenburg;Christine C Genick;K. A. Clark;David A. Schultz
中科院分区：
文献类型：
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作者：
S. Oldenburg;Christine C Genick;K. A. Clark;David A. Schultz
我们展示了使用银等离子共振粒子（PRPs）作为报告标记，在基于微阵列的DNA杂交试验中，我们筛选乳腺癌基因BRCA1中已知的多态性位点。PRPs（直径40-100nm）在配备暗场照明的标准显微镜下作为有色光的衍射极限点成像，可以单独识别和区分背景散射。而不是总强度，在CCD图像中，通过软件算法计算PRPs的数量作为这些分析的信号。在典型的PRP杂交分析中，我们获得的检测灵敏度比使用荧光标记获得的灵敏度高约60倍。我们得出结论，单颗粒计数是稳健的，通常适用于各种分析平台，并且可以集成到低成本和定量检测系统中，用于单核苷酸多态性分析。
We demonstrate the use of silver plasmon resonant particles (PRPs), as reporter labels, in a microarray-based DNA hybridization assay in which we screen for a known polymorphic site in the breast cancer gene BRCA1. PRPs (40–100nm in diameter) image as diffraction-limited points of colored light in a standard microscope equipped with dark-field illumination, and can be individually identified and discriminated against background scatter. Rather than overall intensity, the number of PRPs counted in a CCD image by a software algorithm serves as the signal in these assays. In a typical PRP hybridization assay, we achieve a detection sensitivity that is ∼60× greater than that achieved by using fluorescent labels. We conclude that single particle counting is robust, generally applicable to a wide variety of assay platforms, and can be integrated into low-cost and quantitative detection systems for single nucleotide polymorphism analysis.