Cambridge Graphene Centre, Department of Engineering, University of Cambridge, UK.
Department of Engineering, University of Cambridge, UK.
Nanoscale. 2023 Sep 29;15(37):15304-15317. doi: 10.1039/d3nr01239e.
Identification of non-amplified DNA sequences and single-base mutations is essential for molecular biology and genetic diagnostics. This paper reports a novel sensor consisting of electrochemically-gated graphene coplanar waveguides coupled with a microfluidic channel. Upon exposure to analytes, propagation of electromagnetic waves in the waveguides is modified as a result of interactions with the fringing field and modulation of graphene dynamic conductivity resulting from electrostatic gating. Probe DNA sequences are immobilised on the graphene surface, and the sensor is exposed to DNA sequences which either perfectly match the probe, contain a single-base mismatch or are unrelated. By monitoring the scattering parameters at frequencies between 50 MHz and 50 GHz, unambiguous and reproducible discrimination of the different strands is achieved at concentrations as low as one attomole per litre (1 aM). By controlling and synchronising frequency sweeps, electrochemical gating, and liquid flow in the microfluidic channel, the sensor generates multidimensional datasets. Advanced data analysis techniques are utilised to take full advantage of the richness of the dataset. A classification accuracy >97% between all three sequences is achieved using different Machine Learning models, even in the presence of simulated noise and low signal-to-noise ratios. The sensor exceeds state-of-the-art sensitivity of field-effect transistors and microwave sensors for the identification of single-base mismatches.
鉴定非扩增 DNA 序列和单碱基突变对于分子生物学和遗传诊断至关重要。本文报道了一种新型传感器,由电化学门控石墨烯共面波导与微流道耦合而成。当暴露于分析物时,由于与边缘场的相互作用以及静电门控导致石墨烯动态电导率的调制,波导中电磁波的传播会发生变化。探针 DNA 序列固定在石墨烯表面,传感器暴露于与探针完全匹配、含有单碱基错配或不相关的 DNA 序列。通过在 50MHz 至 50GHz 之间的频率监测散射参数,可以在低至 1 个皮摩尔每升(1aM)的浓度下实现不同链的明确和可重复的区分。通过控制和同步频率扫描、电化学门控以及微流道中的液体流动,传感器生成多维数据集。利用先进的数据分析技术充分利用数据集的丰富性。使用不同的机器学习模型,即使在存在模拟噪声和低信噪比的情况下,也可以实现所有三种序列之间的分类准确率>97%。该传感器在识别单碱基错配方面超过了场效应晶体管和微波传感器的现有技术灵敏度。