Microarray gene expression analysis free of reverse transcription and dye labeling.
Microarray gene expression analysis free of reverse transcription and dye labeling.
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无需逆转录和染料标记的微阵列基因表达分析。
DOI：
10.1016/j.ab.2005.07.023
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发表时间：
2005
影响因子：
2.9
通讯作者：
Golovlev,Val
中科院分区：
文献类型：
--
作者：
Sun,Ye;Fan,Wen-Hua;McCann,MichaelP;Golovlev,Val
使用直径为250nm的阳离子金纳米颗粒作为目标检测试剂，开发了一种用于基因表达分析的新型微阵列系统。该方法利用未标记的目标分子与阵列上的互补探针杂交，然后在胶体金溶液中孵育。由于阳离子金颗粒和目标 DNA 主链中阴离子磷酸基团的静电吸引力，纳米金颗粒在杂交点上沉淀，从而产生杂交信号。与传统的荧光检测相比，这种基于纳米粒子的检测系统消除了目标标记程序。杂交信号的可视化可以用平板扫描仪代替共焦激光扫描仪来完成，这大大简化了过程并降低了成本。灵敏度估计小于阵列表面捕获的 2pg DNA 分子。来自杂交点的信号定量地代表捕获的靶DNA的量，因此允许定量基因表达分析。跨阵列重现性足以检测两个微阵列实验中两倍或更少的信号变化。
A new microarray system has been developed for gene expression analysis using cationic gold nanoparticles with diameters of 250nm as a target detection reagent. The approach utilizes nonlabeled target molecules hybridizing with complementary probes on the array, followed by incubation in a colloidal gold solution. The hybridization signal results from the precipitation of nanogold particles on the hybridized spots due to the electrostatic attraction of the cationic gold particles and the anionic phosphate groups in the target DNA backbone. In contrast to conventional fluorescent detection, this nanoparticle-based detection system eliminates the target labeling procedure. The visualization of hybridization signals can be accomplished with a flatbed scanner instead of a confocal laser scanner, which greatly simplifies the process and reduces the cost. The sensitivity is estimated to be less than 2pg of DNA molecules captured on the array surface. The signal from hybridized spots quantitatively represents the amount of captured target DNA and therefore permits quantitative gene expression analysis. Cross-array reproducibility is adequate for detecting twofold or less signal changes across two microarray experiments.