Laboratory of Process Engineering & Environment, Faculty of Sciences and Techniques, Hassan II, University of Casablanca, B.P.146, Mohammedia 28806, Morocco.
Biosensors (Basel). 2023 Aug 28;13(9):853. doi: 10.3390/bios13090853.
The development of biosensors for target detection plays a crucial role in advancing various fields of bioscience. This work presents the development of a genosensor that exploits the colorimetric phenol-sulfuric acid sugar reaction for the detection of DNA, and RNA as specific targets, and DNA intercalator molecules. The biosensor combines simplicity and reliability to create a novel bioassay for accurate and rapid analysis. A 96-well microplate based on a polystyrene polymer was used as the platform for an unmodified capture DNA immobilization via a silanization process and with (3-Aminopropyl) triethoxysilane (APTES). After that, a hybridization step was carried out to catch the target molecule, followed by adding phenol and sulfuric acid to quantify the amount of DNA or RNA sugar backbone. This reaction generated a yellow-orange color on the wells measured at 490 nm, which was proportional to the target concentration. Under the optimum conditions, a calibration curve was obtained for each target. The developed biosensor demonstrated high sensitivity, good selectivity, and linear response over a wide concentration range for DNA and RNA targets. Additionally, the biosensor was successfully employed for the detection of DNA intercalator agents that inhibited the hybridization of DNA complementary to the immobilized capture DNA. The developed biosensor offers a potential tool for sensitive and selective detection in various applications, including virus diagnosis, genetic analysis, pathogenic bacteria monitoring, and drug discovery.
生物传感器在目标检测方面的发展在推进各个生物科学领域中起着至关重要的作用。本工作提出了一种基于比色酚-硫酸糖反应的基因传感器的开发，用于检测 DNA 和 RNA 作为特定靶标以及 DNA 嵌入剂分子。该生物传感器结合了简单性和可靠性，为准确快速分析创造了一种新的生物测定法。以聚苯乙烯聚合物为基础的 96 孔微孔板被用作平台，通过硅烷化过程和（3-氨丙基）三乙氧基硅烷（APTES）对未修饰的捕获 DNA 进行固定。之后，进行杂交步骤以捕获靶分子，然后加入苯酚和硫酸来定量 DNA 或 RNA 糖主链的量。该反应在 490nm 处测量的孔中生成黄色橙色颜色，与目标浓度成正比。在最佳条件下，为每个目标获得了校准曲线。所开发的生物传感器在 DNA 和 RNA 靶标广泛的浓度范围内表现出高灵敏度、良好的选择性和线性响应。此外，该生物传感器成功用于检测抑制与固定捕获 DNA 互补的 DNA 杂交的 DNA 嵌入剂。所开发的生物传感器为各种应用中的敏感和选择性检测提供了一种潜在的工具，包括病毒诊断、遗传分析、病原细菌监测和药物发现。
