Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Alexandria University, Alexandria 21521, Egypt.
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Alexandria University, Alexandria 21521, Egypt.
Spectrochim Acta A Mol Biomol Spectrosc. 2018 Dec 5;205:48-54. doi: 10.1016/j.saa.2018.07.011. Epub 2018 Jul 7.
A highly selective and sensitive fluorescence assay for kanamycin has been developed that depends on complementation of two splits of DNA aptamer. One DNA split was labeled with CuS nanoparticle and the other was decorated with biotin, which enabled coupling with streptavidin magnesphere paramagnetic particles (PMPs). Complementation of the two-aptamer splits happened only in the presence of kanamycin and the subsequent sandwich was separated via a magnet. The released Cu(II) was reduced to Cu(I) by sodium ascorbate and finally catalyzed the click reaction between fluorogenic 3-azido-7-hydroxycoumarin and propargyl alcohol to afford the corresponding fluorescent 1,4-disubstituted-1,2,3-triazole. The fluorescence signal produced (λ = 365 nm, λ = 470 nm) was dependent on kanamycin concentration. Fluorescence signal amplification was found to be in good linear relationship with the logarithm of kanamycin concentration in the range of 0.04-20 nM. Furthermore, the proposed assay showed a good reproducibility, high selectivity and low detection limits for kanamycin determination. In addition, the capability of the proposed method to detect kanamycin in biological samples with satisfactory results was demonstrated.
已开发出一种高度灵敏和选择性的荧光测定法，用于检测卡那霉素，该方法依赖于两个 DNA 适体片段的互补。一个 DNA 片段用 CuS 纳米粒子标记，另一个片段用生物素修饰，这使得它能够与链霉亲和素磁珠（PMPs）偶联。只有在存在卡那霉素的情况下，两个适体片段才会互补，随后通过磁铁分离夹心结构。释放的 Cu(II)被抗坏血酸钠还原为 Cu(I)，然后催化荧光性 3-叠氮-7-羟基香豆素和丙炔醇之间的点击反应，生成相应的荧光 1,4-二取代-1,2,3-三唑。产生的荧光信号（λ = 365 nm，λ = 470 nm）取决于卡那霉素的浓度。发现荧光信号的放大与卡那霉素浓度的对数在 0.04-20 nM 范围内呈良好的线性关系。此外，该方法还表现出良好的重现性、高选择性和低检测限，可用于卡那霉素的检测。此外，还证明了该方法在生物样品中检测卡那霉素的能力，结果令人满意。
