State Key Laboratory of Marine Resource Utilization in South China Sea, College of materials and chemical engineering, Hainan University, Haikou 570228, China.
Molecules. 2019 Feb 5;24(3):575. doi: 10.3390/molecules24030575.
Simple and rapid detection of DNA single base mismatch or point mutation is of great significance for the diagnosis, treatment, and detection of single nucleotide polymorphism (SNP) in genetic diseases. Homogeneous mutation assays with fast hybridization kinetics and amplified discrimination signals facilitate the automatic detection. Herein we report a quick and cost-effective assay for SNP analysis with a fluorescent single-labeled DNA probe. This convenient strategy is based on the efficient quenching effect and the preferential binding of graphene oxide (GO) to ssDNA over dsDNA. Further, a cationic comb-type copolymer (CCC), poly(l-lysine)--dextran (PLL--Dex), significantly accelerates DNA hybridization and strand-exchange reaction, amplifying the effective distinction of the kinetic barrier between a perfect matched DNA and a mismatched DNA. Moreover, in vitro experiments indicate that RAW 264.7 cells cultured on PLL--Dex exhibits excellent survival and proliferation ability, which makes this mismatch detection strategy highly sensitive and practical.
简单、快速地检测 DNA 单碱基错配或点突变对于遗传疾病的诊断、治疗和单核苷酸多态性（SNP）的检测具有重要意义。具有快速杂交动力学和放大的区分信号的均相突变检测有利于自动检测。本文报道了一种利用荧光单标记 DNA 探针进行 SNP 分析的快速、经济有效的方法。这种方便的策略基于石墨烯氧化物（GO）对 ssDNA 比对 dsDNA 的高效猝灭效应和优先结合。此外，阳离子梳状共聚物（CCC）聚（L-赖氨酸）-葡聚糖（PLL-Dex）显著加速了 DNA 杂交和链交换反应，放大了完美匹配 DNA 和错配 DNA 之间动力学障碍的有效区别。此外，体外实验表明，在 PLL-Dex 上培养的 RAW 264.7 细胞具有优异的存活和增殖能力，这使得这种错配检测策略具有很高的灵敏度和实用性。
