Tang Mingjie, Zhang Mingkun, Xia Liangping, Yang Zhongbo, Yan Shihan, Wang Huabin, Wei Dongshan, Du Chunlei, Cui Hong-Liang
Chongqing Institute of Green and Intelligent Technology, Chinese Academy of Sciences, Chongqing, 400714, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Biomed Opt Express. 2020 Aug 31;11(9):5362-5372. doi: 10.1364/BOE.400487. eCollection 2020 Sep 1.
Many human genetic diseases are caused by single-base mutation in the gene sequence. Since DNA molecules with single-base mutation are extremely difficult to differentiate, existing detection methods are invariably complex and time-consuming. We propose a new label-free and fast terahertz (THz) spectroscopic technique based on a home-made terahertz attenuated total reflection (ATR) microfluidic cell and a terahertz time-domain spectroscopy (THz-TDS) system to detect single-base-mutated DNA molecules. The detected object DNA molecules are normal hemoglobin gene, sickle cell anemia gene (15 nt), JAK2 gene wild type and JAK2 V617F gene mutation (39 nt) from sickle cell anemia and thrombocytopenia, respectively. Results show that the oligonucleotide fragments with single-base mutation can be identified by THz spectroscopy combined with the ATR microfluidic cell, and the recognition effect of short oligonucleotide fragments with single-base mutation is better than that of long oligonucleotide fragments. The terahertz biosensor is shown to have high sensitivity and can be used to detect DNA molecules directly in the solution environment.
许多人类遗传疾病是由基因序列中的单碱基突变引起的。由于单碱基突变的DNA分子极难区分，现有的检测方法总是复杂且耗时的。我们提出了一种基于自制太赫兹衰减全反射（ATR）微流控池和太赫兹时域光谱（THz-TDS）系统的新型无标记快速太赫兹光谱技术，用于检测单碱基突变的DNA分子。检测对象DNA分子分别是来自镰状细胞贫血和血小板减少症的正常血红蛋白基因、镰状细胞贫血基因（15个核苷酸）、JAK2基因野生型和JAK2 V617F基因突变（39个核苷酸）。结果表明，结合ATR微流控池的太赫兹光谱能够识别单碱基突变的寡核苷酸片段，且单碱基突变的短寡核苷酸片段的识别效果优于长寡核苷酸片段。太赫兹生物传感器具有高灵敏度，可用于在溶液环境中直接检测DNA分子。
