Centro de Investigación y Desarrollo Tecnológico en Electroquímica (CIDETEQ), Parque Tecnológico Querétaro s/n, Sanfandila, Pedro Escobedo, C.P. 76703, Querétaro, Qro, Mexico.
Centre for Nanotechnology and Advanced Biomaterials, SASTRA Deemed to be University, Thanjavur, India.
Colloids Surf B Biointerfaces. 2019 Apr 1;176:431-438. doi: 10.1016/j.colsurfb.2019.01.016. Epub 2019 Jan 7.
Seed-mediated Gold-Iron oxide yolk-shell nanoparticles (YSNPs) were synthesized and functionalized with cy5 attached- thiolated single strand DNA probe for the detection of mutated DNA. The optimum concentration of thiolated DNA determined from a bathochromic shift of surface plasmon resonance (SPR) peak, was 0.177μM. The effect of pH (2-10), temperature (4, 37, 60 and 100 °C), and ionic strengths (1 M to 4 M) on the stability of ssDNA probe tethered YSNPs, studied with the assistance of flocculation parameter. The detection of mutation in DNA was possible using such ssDNA probe functionalized and stabilized nanoparticles. The hybridization of the oligonucleotide probe with the complementary, non-complementary and mutated DNA strands are determined via their respective intensities of the fluorescence of cy5, an efficient fluorescent marker. The intensities help in the comprehension of the specificity of the system. The report predicts controlled efficiency of hybridization with the aid of Hamaker constant, which is determined as 1.15 × 10 J for DNA functionalized YSNPs. The minimum concentration of target DNA detected using this methodology was 1.2 × 10 mol/L.
采用种子介导法合成了金-氧化铁蛋黄壳纳米粒子（YSNPs），并用连接有 Cy5 的巯基化单链 DNA 探针对其进行功能化，用于检测突变 DNA。通过表面等离子体共振（SPR）峰的红移确定了最佳的巯基化 DNA 浓度为 0.177μM。利用絮凝参数研究了 pH 值（2-10）、温度（4、37、60 和 100°C）和离子强度（1 M 至 4 M）对 ssDNA 探针固定化 YSNPs 稳定性的影响。使用这种 ssDNA 探针功能化和稳定的纳米粒子可以检测 DNA 中的突变。通过寡核苷酸探针与互补、非互补和突变 DNA 链的杂交，可以通过 Cy5 的荧光强度来确定，Cy5 是一种有效的荧光标记物。这些强度有助于理解该系统的特异性。报告预测，通过 Hamaker 常数可以控制杂交效率，对于 DNA 功能化的 YSNPs，Hamaker 常数确定为 1.15×10^-19 J。使用这种方法检测到的最小靶 DNA 浓度为 1.2×10^-10 mol/L。
