Department of Chemistry, University of Isfahan, Isfahan 81746-73441, Iran.
Department of Chemistry, University of Isfahan, Isfahan 81746-73441, Iran.
Bioelectrochemistry. 2017 Oct;117:83-88. doi: 10.1016/j.bioelechem.2017.06.006. Epub 2017 Jun 15.
In the present study, a nanoporous gold platform was applied for the amplified detection of Hepatitis B virus (HBV) by an electrochemical DNA biosensor. Ferrocene as a redox reporter was covalently attached to the DNA probe and its electrochemical signal was recorded as the biosensor response. For real samples, DNA was firstly extracted from blood of patients and then amplified by polymerase chain reaction (PCR) for 5cycles. Sensitivity of this biosensor was enhanced by using nanoporous gold electrode, therefore this sensor can discriminate the genome of HBV in real sample with low PCR cycles. By this strategy and signal amplification using nanoporous platform and covalently attached electroactive label, the biosensor can distinguish between healthy and HBV patients with limited PCR cycles. Moreover, the errors of PCR with large cycles can be disregarded. A linear dynamic range of 0.4 to 10nmol of mutant DNA was achieved, with reliable reproducibility (RSD) 8.9%.
在本研究中，通过电化学 DNA 生物传感器应用纳米多孔金平台对乙型肝炎病毒 (HBV) 进行了放大检测。二茂铁作为一种氧化还原报告分子被共价连接到 DNA 探针上，并记录其电化学信号作为生物传感器的响应。对于实际样品，首先从患者的血液中提取 DNA，然后通过聚合酶链反应 (PCR) 扩增 5 个循环。通过使用纳米多孔金电极，提高了生物传感器的灵敏度，因此该传感器可以在低 PCR 循环数下区分实际样本中的 HBV 基因组。通过这种策略和使用纳米多孔平台和共价附着的电活性标记进行信号放大，该传感器可以区分健康人和 HBV 患者，即使 PCR 循环数有限。此外，可以忽略 PCR 大循环数带来的误差。该方法实现了突变型 DNA 的线性动态范围为 0.4 至 10nmol，具有可靠的重现性（RSD）为 8.9%。
