Dipartimento di Chimica, Università di Firenze, Sesto Fiorentino, Italy.
Anal Bioanal Chem. 2013 Jan;405(2-3):985-93. doi: 10.1007/s00216-012-6345-4. Epub 2012 Sep 6.
The work presented here deals with the optimization of a strategy for detection of single nucleotide polymorphisms based on surface plasmon resonance imaging. First, a sandwich-like assay was designed, and oligonucleotide sequences were computationally selected in order to study optimized conditions for the detection of the rs1045642 single nucleotide polymorphism in the gene ABCB1. Then the strategy was optimized on a surface plasmon resonance imaging biosensor using synthetic DNA sequences in order to evaluate the best conditions for the detection of a single mismatching base. Finally, the assay was tested on DNA extracted from human blood which was subsequently amplified using a whole genome amplification kit. The direct detection of the polymorphism was successfully achieved. The biochip was highly regenerable and reusable for up to 20 measurements. Furthermore, coupling these promising results with the multiarray assay, we can foresee applying this biosensor in clinical research extended to concurrent analysis of different polymorphisms.
这里介绍的工作涉及基于表面等离子体共振成像的单核苷酸多态性检测策略的优化。首先，设计了一种夹心样测定法，并通过计算选择了寡核苷酸序列，以便研究 ABCB1 基因中 rs1045642 单核苷酸多态性的最佳检测条件。然后，使用合成 DNA 序列在表面等离子体共振成像生物传感器上对该策略进行了优化，以评估检测单个错配碱基的最佳条件。最后，在使用全基因组扩增试剂盒扩增的人血 DNA 上测试了该测定法。成功实现了对多态性的直接检测。生物芯片具有高度的可再生性和可重复使用性，可进行多达 20 次测量。此外，将这些有前途的结果与多阵列测定法结合使用，我们可以预见将这种生物传感器应用于临床研究，扩展到同时分析不同的多态性。
