Zhu Debin, Tang Yabing, Xing Da, Chen Wei R
MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University, Guangzhou 510631, China.
Anal Chem. 2008 May 15;80(10):3566-71. doi: 10.1021/ac0713306. Epub 2008 Apr 3.
A bio bar code assay based on oligonucleotide-modified gold nanoparticles (Au-NPs) provides a PCR-free method for quantitative detection of nucleic acid targets. However, the current bio bar code assay requires lengthy experimental procedures including the preparation and release of bar code DNA probes from the target-nanoparticle complex and immobilization and hybridization of the probes for quantification. Herein, we report a novel PCR-free electrochemiluminescence (ECL)-based bio bar code assay for the quantitative detection of genetically modified organism (GMO) from raw materials. It consists of tris-(2,2'-bipyridyl) ruthenium (TBR)-labeled bar code DNA, nucleic acid hybridization using Au-NPs and biotin-labeled probes, and selective capture of the hybridization complex by streptavidin-coated paramagnetic beads. The detection of target DNA is realized by direct measurement of ECL emission of TBR. It can quantitatively detect target nucleic acids with high speed and sensitivity. This method can be used to quantitatively detect GMO fragments from real GMO products.
基于寡核苷酸修饰金纳米颗粒（Au-NPs）的生物条形码分析提供了一种无需聚合酶链式反应（PCR）的核酸靶标定量检测方法。然而，当前的生物条形码分析需要冗长的实验步骤，包括从靶标-纳米颗粒复合物中制备和释放条形码DNA探针，以及将探针固定和杂交以进行定量分析。在此，我们报告了一种新型的基于无PCR电化学发光（ECL）的生物条形码分析方法，用于从原材料中定量检测转基因生物（GMO）。它由三（2,2'-联吡啶）钌（TBR）标记的条形码DNA、使用Au-NPs和生物素标记探针的核酸杂交，以及通过链霉亲和素包被的顺磁性珠子选择性捕获杂交复合物组成。通过直接测量TBR的ECL发射来实现靶标DNA的检测。它能够以高速和高灵敏度定量检测靶标核酸。该方法可用于从实际转基因产品中定量检测转基因片段。
