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基于单线态氧的光电化学 DNA 检测。

Singlet oxygen-based photoelectrochemical detection of DNA.

机构信息

A-Sense Lab, Department of Bioengineering, University of Antwerp, Groenenborgerlaan 171, 2020, Antwerp, Belgium; NANOlab Center of Excellence, University of Antwerp, Groenenborgerlaan 171, 2020, Antwerp, Belgium.

A-Sense Lab, Department of Bioengineering, University of Antwerp, Groenenborgerlaan 171, 2020, Antwerp, Belgium; NANOlab Center of Excellence, University of Antwerp, Groenenborgerlaan 171, 2020, Antwerp, Belgium.

出版信息

Biosens Bioelectron. 2022 Jan 1;195:113652. doi: 10.1016/j.bios.2021.113652. Epub 2021 Sep 23.

Abstract

The current work, designed for the photoelectrochemical detection of DNA, evaluates light-responsive DNA probes carrying molecular photosensitizers generating singlet oxygen (O). We take advantage of their chromophore's ability to produce O upon photoexcitation and subsequent photocurrent response. Type I, fluorescent and type II photosensitizers were studied using diode lasers at 406 nm blue, 532 nm green and 659 nm red lasers in the presensce and absence of a redox reporter, hydroquinone (HQ). Only type II photosensitizers (producing O) resulted in a noticeable photocurrent in 1-4 nA range upon illumination, in particular, dissolved DNA probes labeled with chlorin e6 and erythrosine were found to give a well-detectable photocurrent response in the presence of HQ. Whereas, Type I photosensitizers and fluorescent chromophores generate negligible photocurrents (<0.15 nA). The analytical performance of the sensing system was evaluated using a magnetic beads-based DNA assay on disposable electrode platforms, with a focus to enhance the sensitivity and robustness of the technique in detecting complementary DNA targets. Amplified photocurrent responses in the range of 70-100 nA were obtained and detection limits of 17 pM and 10 pM were achieved using magnetic beads-captured chlorin e6 and erythrosine labeled DNA probes respectively. The presented novel photoelectrochemical detection can further be optimized and employed in applications for which enzymatic amplification such as polymerase chain reaction (PCR) is not applicable owing to their limitations and as an effective alternative to colorimetric detection when rapid detection of specific nucleic acid targets is required.

摘要

当前的工作旨在设计用于光电化学检测 DNA 的光响应 DNA 探针,这些探针携带分子光敏剂,可产生单线态氧 (O)。我们利用发色团在光激发后产生 O 和随后的光电流响应的能力。使用二极管激光器在 406nm 蓝光、532nm 绿光和 659nm 红光下研究了 I 型、荧光和 II 型光敏剂,在存在和不存在氧化还原报告分子对苯二酚 (HQ) 的情况下。只有 II 型光敏剂(产生 O)在光照下会产生明显的光电流,范围在 1-4nA 之间,特别是用氯卟啉 e6 和赤藓红标记的溶解 DNA 探针在存在 HQ 的情况下会产生可检测的光电流响应。相比之下,I 型光敏剂和荧光发色团产生的光电流很小(<0.15nA)。通过在一次性电极平台上基于磁珠的 DNA 测定评估了传感系统的分析性能,重点是提高技术检测互补 DNA 靶标的灵敏度和稳健性。使用磁珠捕获的氯卟啉 e6 和赤藓红标记的 DNA 探针分别获得了 70-100nA 的放大光电流响应,检测限分别达到了 17pM 和 10pM。提出的新型光电化学检测方法可以进一步优化,并在需要快速检测特定核酸靶标而酶扩增(如聚合酶链反应 (PCR))不适用的情况下应用,也可以作为比色检测的有效替代方法。

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