Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources, Ministry of Education, College of Chemistry and Chemical Engineering, Guangxi Normal University, Guilin 541004, PR China.
Biosens Bioelectron. 2013 Apr 15;42:598-602. doi: 10.1016/j.bios.2012.11.025. Epub 2012 Nov 27.
An amplified graphene oxide (GO) based fluorescence aptasensor based on target-triggered aptamer hairpin switch and strand-displacement polymerization recycling is developed for bioassays. The dye-labeled single-strand DNA (aptamer hairpin) was adsorbed on the surface of GO, which result in the fluorescence quenching of dye, and exhibiting minimal background fluorescence. Upon the target, primer and polymerase, the stem of the aptamer hairpin was opened, and binds with the primer to triggers the circular target strand-displacement polymerization reaction, which produces huge amounts of duplex helixes DNA and lead to strong fluorescence emission due to shielding of nucelobases within its double-helix structure. During the polymerization reaction, the primer was extended, and target was displaced. And the displaced target recognizes and hybridizes with another hairpin probe, triggering the next round of polymerization reaction, and the circle process induces fluorescence signal amplification for the detection of analyte. To test the feasibility of the aptasensor systems, interferon-gamma (IFN-γ) was employed as a model analyte. A detection limit as low as 1.5 fM is obtained based on the GO aptasensor with a linear range of three orders of magnitude. The present method was successfully applied for the detection of IFN-γ in human plasma.
基于靶标触发的适体发夹开关和链置换聚合循环的放大氧化石墨烯(GO)基荧光适体传感器用于生物分析。标记染料的单链 DNA(适体发夹)被吸附在 GO 表面,导致染料的荧光猝灭,并表现出最小的背景荧光。在靶标、引物和聚合酶的作用下,适体发夹的茎部被打开,并与引物结合,触发环状靶标链置换聚合反应,产生大量双链 DNA,并由于其双螺旋结构内的核碱基被屏蔽而导致强荧光发射。在聚合反应过程中,引物被延伸,靶标被置换。被置换的靶标识别并与另一个发夹探针杂交,触发下一轮聚合反应,该循环过程诱导荧光信号放大,用于分析物的检测。为了测试适体传感器系统的可行性,将干扰素-γ(IFN-γ)用作模型分析物。基于 GO 适体传感器,检测限低至 1.5 fM,线性范围为三个数量级。该方法成功地用于人血浆中 IFN-γ 的检测。