Department of Chemical and Environmental Engineering, University of California, Riverside, California 92521-9800, United States.
Department of Industrial & Systems Engineering, Rochester Institute of Technology, Rochester, New York 14623-5603, United States.
Anal Chem. 2023 Aug 15;95(32):11997-12005. doi: 10.1021/acs.analchem.3c01724. Epub 2023 Jul 28.
An aerosol jet printing-enabled dual-function biosensor for the sensitive detection of pathogens using SARS-CoV-2 RNA as an example has been developed. A CRISPR-Cas13:guide-RNA complex is activated in the presence of a target RNA, leading to the collateral trans-cleavage of ssRNA probes that contain a horseradish peroxidase (HRP) tag. This, in turn, catalyzes the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by HRP, resulting in a color change and electrochemical signal change. The colorimetric and electrochemical sensing protocol does not require complicated target amplification and probe immobilization and exhibits a detection sensitivity in the femtomolar range. Additionally, our biosensor demonstrates a wide dynamic range of 5 orders of magnitude. This low-cost aerosol inkjet printing technique allows for an amplification-free and integrated dual-function biosensor platform, which operates at physiological temperature and is designed for simple, rapid, and accurate point-of-care (POC) diagnostics in either low-resource settings or hospitals.
已开发出一种气溶胶喷射打印功能的双功能生物传感器,以 SARS-CoV-2 RNA 为例,可实现对病原体的灵敏检测。在存在靶 RNA 的情况下,CRISPR-Cas13:guide-RNA 复合物被激活,导致含有辣根过氧化物酶 (HRP) 标签的 ssRNA 探针的旁切裂解。这反过来又催化 HRP 氧化 3,3',5,5'-四甲基联苯胺 (TMB),导致颜色和电化学信号变化。比色和电化学生感协议不需要复杂的目标扩增和探针固定,并表现出飞摩尔范围内的检测灵敏度。此外,我们的生物传感器还表现出 5 个数量级的宽动态范围。这种低成本的气溶胶喷墨打印技术允许无扩增和集成的双功能生物传感器平台,该平台在生理温度下运行,专为在低资源环境或医院中进行简单、快速和准确的即时护理 (POC) 诊断而设计。