Professur für Physikalische Chemie, Mess- und Sensortechnik, Technische Universität Dresden, 01062 Dresden, Germany.
Biosens Bioelectron. 2010 Dec 15;26(4):1543-7. doi: 10.1016/j.bios.2010.07.108. Epub 2010 Aug 4.
We report on the development of a new platform technology for the detection of genetic variations by means of surface plasmon resonance (SPR) spectroscopy. TOPAS chips with integrated optics were exploited in combination with microfluidics. Within minutes, the detection of hybridization kinetics was achieved simultaneously at all spots of the DNA microarray. A nanoliter dispenser is used to deposit thiol-modified single-stranded probe DNA on the gold surface of the chips. We investigated the influence of different parameters on hybridization using model polymerase chain reaction (PCR) products. These PCR products comprised a single-stranded tag sequence being complementary to an anti-tag sequence of probes immobilized on the gold surface. The signals increased with increasing length of PCR products (60, 100 or 300 base pairs) as well as with their concentration. We investigated hybridizations on DNA microarrays comprising 90 spots of probe DNA with three different sequences. Furthermore, we demonstrate that sequences with possible hairpin structures significantly lower the binding rate, and thus, the SPR signals during hybridization.
我们报告了一种新的平台技术的发展，用于通过表面等离子体共振（SPR）光谱检测遗传变异。TOPAS 芯片与集成光学相结合，结合微流控技术使用。在几分钟内，DNA 微阵列的所有点的杂交动力学检测都可以同时实现。纳米升分配器用于将巯基修饰的单链探针 DNA 沉积在芯片的金表面上。我们使用模型聚合酶链反应（PCR）产物研究了不同参数对杂交的影响。这些 PCR 产物包含与固定在金表面上的探针的反标签序列互补的单链标签序列。信号随着 PCR 产物（60、100 或 300 个碱基对）的长度以及浓度的增加而增加。我们研究了包含 90 个探针 DNA 点的 DNA 微阵列上的杂交，这些探针 DNA 具有三个不同的序列。此外，我们证明了具有可能发夹结构的序列会显着降低结合率，从而降低杂交过程中的 SPR 信号。
