Department of Microbiology, Hunan Normal University, Changsha, Hunan, 410081, People's Republic of China.
J Fluoresc. 2010 Jan;20(1):365-9. doi: 10.1007/s10895-009-0513-8. Epub 2009 Sep 26.
Fluorescence resonance energy transfer (FRET) that consists of quantum dot as donors and organic fluorophore dyes as acceptors has been a very important method to detect biomolecules such as nucleic acids. In this work, we established a new FRET detection system of Bifidobacterium species-specific 16S rDNA using QD-ROX FRET bioprobe, in which 525 nm QD-DNA conjugation consisted of the carboxyl-modified QD and the amino-modified DNA in the presence of EDC. Both ROX-DNA and the conjugation above could hybridize with the target DNA after forming the QD-ROX bioprobe. When the hybridization made the distance between the QD and ROX to meet FRET effect needed, 525 nm QD fluorescence intensity decreased and ROX fluorescence intensity increased. In the control, there was no notable change of fluorescence intensities without target DNA. It is very clear that the change of the QD and ROX fluorescence intensities provide the good base and guaranty for this rapid and simple detection system.
荧光共振能量转移(FRET)由量子点作为供体和有机荧光染料作为受体组成,是检测生物分子如核酸的一种非常重要的方法。在这项工作中,我们建立了一种使用 QD-ROX FRET 生物探针检测双歧杆菌属特异性 16S rDNA 的新 FRET 检测系统,其中 525nm 的 QD-DNA 缀合物由在 EDC 存在下羧基修饰的 QD 和氨基修饰的 DNA 组成。ROX-DNA 和上述缀合物都可以与目标 DNA 杂交形成 QD-ROX 生物探针。当杂交使 QD 和 ROX 之间的距离达到 FRET 效应所需的距离时,525nm 的 QD 荧光强度降低,ROX 荧光强度增加。在对照实验中,没有目标 DNA 时,荧光强度没有明显变化。很明显,QD 和 ROX 荧光强度的变化为这种快速简单的检测系统提供了良好的基础和保证。