Department of Mechanical Engineering, Korea University, Seoul, 02841, Republic of Korea.
School of Mechanical Engineering, Hoseo University, Asan, 31499, Republic of Korea.
Talanta. 2019 Dec 1;205:120154. doi: 10.1016/j.talanta.2019.120154. Epub 2019 Jul 16.
Mutant DNAs are important markers useful for the diagnosis of human disease. Single-nucleotide polymorphisms (SNPs) represent the most common types of DNA mutations. As there is only a one base pair change in a single nucleotide between the SNP and the wild-type DNA, it is difficult to distinguish the SNPs. In this report, a highly sensitive and selective detection and discrimination of SNPs is performed using MutS, gold nanoparticles (AuNP) and a resonator. A single mismatched base exists between the SNP mutation and the probe DNA on the resonator, and MutS binds to the DNA at the location of the mismatch. As MutS is attached to AuNP (MutS-AuNP), both MutS and AuNP are adsorbed onto the resonator. The detection is based on the resonance frequency shift of the resonator following the adsorption of MutS-AuNP on the resonator. Highly sensitive detection of DNA mutations was achieved using AuNPs that act as mass amplifiers, and the obtained limit of detected was 100 fM. Additionally, our proposed method detected mutations in the presence of as little as 0.1% wild-type, and discrimination of specific mutations was also achieved. The results obtained from our proposed method suggest its potential for diagnosing cancer patients.
突变 DNA 是用于人类疾病诊断的重要标志物。单核苷酸多态性 (SNP) 是最常见的 DNA 突变类型。由于 SNP 和野生型 DNA 之间只有一个碱基对的变化，因此很难区分 SNP。在本报告中，使用 MutS、金纳米粒子 (AuNP) 和共振器对 SNP 进行了高灵敏度和选择性的检测和区分。在共振器上的 SNP 突变和探针 DNA 之间存在单个错配碱基，MutS 结合到错配位置的 DNA 上。由于 MutS 附着在 AuNP（MutS-AuNP）上，因此 MutS 和 AuNP 都被吸附到共振器上。检测是基于 MutS-AuNP 吸附到共振器上后共振器的共振频率位移。使用作为质量放大器的 AuNP 实现了对 DNA 突变的高灵敏度检测，并且获得的检测限为 100 fM。此外，我们提出的方法在存在低至 0.1%野生型的情况下也可以检测到突变，并实现了对特定突变的区分。我们提出的方法的结果表明其在诊断癌症患者方面具有潜力。
