Aslan Kadir, Malyn Stuart N, Bector Geetika, Geddes Chris D
Institute of Fluorescence, Laboratory for Advanced Medical Plasmonics, Medical Biotechnology Center, University of Maryland Biotechnology Institute, Baltimore, MD 21201, USA.
Analyst. 2007 Nov;132(11):1122-9. doi: 10.1039/b708069g. Epub 2007 Sep 11.
In this paper, we investigated the effects of low-power microwave heating on the components of the recently described new approach to surface DNA hybridization assays, based on the Microwave-Accelerated Metal-Enhanced Fluorescence (MAMEF) platform technology. Thiolated oligonucleotides have been linked to surface-bound silver nanostructures which partially coat a glass slide. The addition of a complementary fluorescein-labeled oligonucleotide results in metal-enhanced fluorescein emission as the probe is brought into close proximity to the silver upon hybridization. In addition, the combined use with low-power microwave heating, which is thought to locally heat around the silvered surface, affords for both the assay kinetics and optical amplification to also be localized to the surface. In our model DNA target assay reported here, we can detect 23-mer targets in less than 20 s, up to a 600-fold decrease in the assay run time as compared to control samples hybridized to completion at room temperature. Importantly, the use of MAMEF also reduces the extent of unwanted non-specific DNA absorption, further increasing specific DNA target detection limits. It was also found that low-power microwave heating did not denature DNA and the bulk temperature increase near to silver nanoparticles was only ca. 1 degrees C.
在本文中,我们研究了基于微波加速金属增强荧光(MAMEF)平台技术的低功率微波加热对最近描述的表面DNA杂交检测新方法的组分的影响。硫醇化寡核苷酸已与部分覆盖载玻片的表面结合的银纳米结构相连。当加入互补的荧光素标记寡核苷酸时,由于杂交时探针与银紧密靠近,会产生金属增强的荧光素发射。此外,与低功率微波加热联合使用,据认为其可局部加热镀银表面周围区域,使得检测动力学和光学放大作用也都局限于表面。在我们在此报道的模型DNA靶标检测中,我们能够在不到20秒内检测到23聚体靶标,与在室温下杂交至完成的对照样品相比,检测运行时间最多减少了600倍。重要的是,MAMEF的使用还降低了不必要的非特异性DNA吸附程度,进一步提高了特异性DNA靶标的检测限。还发现低功率微波加热不会使DNA变性,银纳米颗粒附近的整体温度升高仅约1℃。