Shanghai Key Laboratory of Chemical Assessment and Sustainability, School of Chemical Science and Engineering, College of Environmental Science and Technology, Tongji University, 1239 Siping Road, Shanghai, 200092, China.
China National Tobacco Quality Supervision & Test Center, Zhengzhou, Henan, 450001, China.
Anal Bioanal Chem. 2017 Nov;409(29):6821-6829. doi: 10.1007/s00216-017-0641-y. Epub 2017 Sep 29.
Detection of DNA damage caused by ∙OH or radiation has led to rapidly growing interest in the fields of drug development, biochemistry, clinic diagnostics, and environmental evaluation. Electrochemical methods have been applied for DNA damage detection because of their fast and sensitive response. However, most of the electrochemical methods for DNA damage detection commonly require immobilization of the strands on the electrode surface. In the present work, sensitive electrochemical monitoring of DNA damage was realized successfully on the basis of in situ DNA chain growth by use of a hybridization chain reaction (HCR) technique. By use of [Ru(NH)] as the signal probe and the Fenton reaction as the ∙OH generator, ultrasensitive detection of DNA damage induced by ∙OH was realized successfully through differential pulse voltammetry with a linear relationship of ∙OH concentration from 15 to 750 pM and a detection limit of 12 pM. Furthermore, environmental DNA-damaging UV light was tested as the lesion source to demonstrate the practicability and reliability of the proposed HCR-based amplified signal method for DNA damage detection. By integrating the HCR technique with an electrochemical method, we provide a promising alternative approach to extend the applications of electrochemical methods in bioanalytical detection of DNA damage. Graphical abstract Sensitive electrochemical monitoring of DNA damage has been realized successfully on the basis of in situ DNA chain growth by use of hybridization chain reaction (HCR). Detection of DNA damage caused by ∙OH was realized successfully through differential pulse voltammetry with a linear relationship of ∙OH concentration from 15 to 750 pM and a detection limit of 12 pM.
·OH 或辐射引起的 DNA 损伤的检测，使得药物开发、生物化学、临床诊断和环境评估等领域对其产生了浓厚的兴趣。电化学方法因其快速灵敏的响应而被应用于 DNA 损伤检测。然而，大多数电化学方法通常需要将链固定在电极表面上来检测 DNA 损伤。在本工作中，基于杂交链式反应（HCR）技术的原位 DNA 链延伸，成功实现了对 DNA 损伤的灵敏电化学监测。利用[Ru（NH3）6]3+作为信号探针，芬顿反应作为·OH 发生器，通过差分脉冲伏安法实现了对·OH 诱导的 DNA 损伤的超灵敏检测，其·OH 浓度在 15 至 750 pM 范围内呈线性关系，检测限低至 12 pM。此外，以环境 DNA 损伤性紫外光作为损伤源，验证了所提出的基于 HCR 的放大信号方法用于 DNA 损伤检测的实用性和可靠性。通过将 HCR 技术与电化学方法相结合，为电化学方法在生物分析检测 DNA 损伤中的应用提供了一种很有前途的替代方法。
