Cheng Cheng, Oueslati Rania, Wu Jayne, Chen Jiangang, Eda Shigetoshi
Department of Electrical Engineering and Computer Science, The University of Tennessee, Knoxville, TN, USA.
Department of Public Health, The University of Tennessee, Knoxville, TN, USA.
Electrophoresis. 2017 Jun;38(12):1617-1623. doi: 10.1002/elps.201700043. Epub 2017 Apr 20.
This work presents a rapid, highly sensitive, low-cost, and specific capacitive DNA sensor for detection of whole genome human herpesvirus-1 DNA. This sensor is capable of direct DNA detection with a response time of 30 s, and it can be used to test standard buffer or serum samples. The sensing approach for DNA detection is based on alternating current (AC) electrokinetics. By applying an inhomogeneous AC electric field on sensor electrodes, positive dielectrophoresis is induced to accelerate DNA hybridization. The same applied AC signal also directly measures the hybridization of target with the probe on the sensor surface. Experiments are conducted to optimize the AC signal, as well as the buffers for probe immobilization and target DNA hybridization. The assay is highly sensitive and specific, with no response to human herpesvirus-2 DNA at 5 ng/mL and a LOD of 1.0 pg/mL (6.5 copies/μL or 10.7 aM) in standard buffer. When testing the double stranded (ds) DNA spiked in human serum samples, the sensor yields a LOD of 20.0 pg/mL (129.5 copies/μL or 0.21 femtomolar (fM)) in neat serum. In this work, the target is whole genome dsDNA, consequently the test can be performed without the use of enzyme or amplification, which considerably simplifies the sensor operation and is highly suitable for point of care disease diagnosis.
这项工作展示了一种用于检测全基因组人类疱疹病毒1型DNA的快速、高灵敏度、低成本且特异的电容式DNA传感器。该传感器能够直接检测DNA,响应时间为30秒,可用于检测标准缓冲液或血清样本。DNA检测的传感方法基于交流(AC)电动学。通过在传感器电极上施加非均匀交流电场,诱导正介电泳以加速DNA杂交。施加的相同交流信号还可直接测量传感器表面目标与探针的杂交情况。进行实验以优化交流信号以及用于探针固定和目标DNA杂交的缓冲液。该检测方法高度灵敏且特异,在标准缓冲液中对5 ng/mL的人类疱疹病毒2型DNA无响应,检测限为1.0 pg/mL(6.5拷贝/μL或10.7 aM)。在检测加标于人类血清样本中的双链(ds)DNA时,该传感器在纯血清中的检测限为20.0 pg/mL(129.5拷贝/μL或0.21飞摩尔(fM))。在这项工作中,目标是全基因组dsDNA,因此无需使用酶或扩增即可进行检测,这大大简化了传感器操作,非常适合即时护理疾病诊断。