Misra Arvind, Shahid Mohammad
Department of Chemistry, Banaras Hindu University, Varanasi, India.
Bioorg Med Chem. 2009 Aug 15;17(16):5826-33. doi: 10.1016/j.bmc.2009.07.015. Epub 2009 Jul 24.
A new sensitive method for the detection of nucleic acids on a glass surface has been described. The self-quenched DNA hairpin probe is immobilized on a glass surface utilizing heterobifunctional reagent, N-(3-triethoxysilylpropyl)-4-(isothiocyanatomethyl)-cyclohexane-1-carboxamide (TPICC). In the closed state fluorescence intensity was quenched due to the presence of guanosine residues in close vicinity of fluorophore while on hybridization with perfectly matched complementary target strand fluorescence was restored. Efficiency and specificity of immobilization as well as thermal stability at variable temperature and pH conditions have been discussed in detail. The method employed has potential for the detection of single nucleotide variations and other diagnostic studies.
本文描述了一种用于检测玻璃表面核酸的新的灵敏方法。利用异双功能试剂N-(3-三乙氧基甲硅烷基丙基)-4-(异硫氰酸酯甲基)-环己烷-1-甲酰胺(TPICC)，将自猝灭DNA发夹探针固定在玻璃表面。在封闭状态下，由于荧光团附近存在鸟苷残基，荧光强度被猝灭，而与完全匹配的互补靶链杂交时，荧光得以恢复。详细讨论了固定化的效率和特异性，以及在不同温度和pH条件下的热稳定性。所采用的方法具有检测单核苷酸变异和其他诊断研究的潜力。
