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A 96-Well LED Array for Multiplexed Photoelectrochemical Detection of Nucleic Acids
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-09-12 , DOI: 10.1021/acs.analchem.4c01998 Saranya Thiruvottriyur Shanmugam 1, 2 , Annemarijn Steijlen 1, 2 , Dennis Laurijssen 3 , Rui Campos 1, 2 , Jan Steckel 3 , Walter Daems 3 , Simone Bassini 1, 2 , Elise Daems 1, 2 , Karolien De Wael 1, 2
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-09-12 , DOI: 10.1021/acs.analchem.4c01998 Saranya Thiruvottriyur Shanmugam 1, 2 , Annemarijn Steijlen 1, 2 , Dennis Laurijssen 3 , Rui Campos 1, 2 , Jan Steckel 3 , Walter Daems 3 , Simone Bassini 1, 2 , Elise Daems 1, 2 , Karolien De Wael 1, 2
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Photoelectrochemical detection of nucleic acid-based cancer biomarkers offers opportunities for highly sensitive, selective, and fast quantitative detection using low-cost measurement instruments. In order to establish itself as a standard method for identifying and quantifying nucleic acids, we have developed a multiplexing strategy using LED technology for photoelectrochemical detection in 96 samples simultaneously. A dedicated setup based on the 96-well plate configuration with a custom-made 96-well LED array was developed. Subsequently, a proof-of-concept study was performed for three miRNAs that are associated with prostate cancer, i.e., miRNA-141, miRNA-145, and miRNA-375. First, measurements with photosensitizer chlorin e6 and redox reporter hydroquinone free in solution proved the proper functioning of the multiplexed detection. Second, the photoelectrochemical detection of the three miRNAs at 24 nM levels was successfully demonstrated. Thereafter, linear calibration curves (R2 > 0.9 for all analytes) were made with plasma spiked with 8–500 pM miRNA. This work presents the first system for multiplexed high-throughput photoelectrochemical detection, allowing it potentially to become a cost-effective and faster alternative to RT-qPCR and gene sequencing techniques in the future.
中文翻译：
用于核酸多重光电化学检测的 96 孔 LED 阵列
基于核酸的癌症生物标志物的光电化学检测为使用低成本测量仪器进行高灵敏度、选择性和快速定量检测提供了机会。为了将其确立为鉴定和定量核酸的标准方法，我们开发了一种使用 LED 技术同时对 96 个样品进行光电化学检测的多重检测策略。开发了基于 96 孔板配置和定制 96 孔 LED 阵列的专用装置。随后，对与前列腺癌相关的三种 miRNA（即 miRNA-141、miRNA-145 和 miRNA-375）进行了概念验证研究。首先，使用溶液中不含光敏剂二氢卟酚 e6 和氧化还原报告剂氢醌的测量证明了多重检测的正确功能。其次，成功证明了 24 nM 水平的三种 miRNA 的光电化学检测。此后，用掺有 8-500 pM miRNA 的血浆制作线性校准曲线（所有分析物的R 2 > 0.9）。这项工作提出了第一个用于多重高通量光电化学检测的系统，使其有可能在未来成为 RT-qPCR 和基因测序技术的一种经济高效、速度更快的替代方案。
更新日期：2024-09-12
中文翻译：
用于核酸多重光电化学检测的 96 孔 LED 阵列
基于核酸的癌症生物标志物的光电化学检测为使用低成本测量仪器进行高灵敏度、选择性和快速定量检测提供了机会。为了将其确立为鉴定和定量核酸的标准方法，我们开发了一种使用 LED 技术同时对 96 个样品进行光电化学检测的多重检测策略。开发了基于 96 孔板配置和定制 96 孔 LED 阵列的专用装置。随后，对与前列腺癌相关的三种 miRNA（即 miRNA-141、miRNA-145 和 miRNA-375）进行了概念验证研究。首先，使用溶液中不含光敏剂二氢卟酚 e6 和氧化还原报告剂氢醌的测量证明了多重检测的正确功能。其次，成功证明了 24 nM 水平的三种 miRNA 的光电化学检测。此后，用掺有 8-500 pM miRNA 的血浆制作线性校准曲线（所有分析物的R 2 > 0.9）。这项工作提出了第一个用于多重高通量光电化学检测的系统，使其有可能在未来成为 RT-qPCR 和基因测序技术的一种经济高效、速度更快的替代方案。
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