生物传感器
环介导等温扩增
核酸
结核分枝杆菌复合物
多重位移放大
比色法
适体
结核分枝杆菌
检出限
DNA
清脆的
化学
聚合酶链反应
生物
分子生物学
材料科学
纳米技术
肺结核
色谱法
基因
生物化学
DNA提取
病理
医学
作者
Mengqi Chen,Xue Jiang,Qianfang Hu,Jinyan Long,Jianwei He,Yuchen Wu,Zhili Wu,Yanhong Niu,Chunmei Jing,Xiaolan Yang
出处
期刊:ACS Sensors
[American Chemical Society]
日期:2023-12-21
卷期号:9 (1): 62-72
被引量:3
标识
DOI:10.1021/acssensors.3c01622
摘要
Rapid and simple nucleic acid detection is significant for disease diagnosis and pathogen screening, especially under specific conditions. However, achieving highly sensitive and specific nucleic acid detection to meet the time and equipment demand remains technologically challenging. In this study, we proposed a magnetic separation enhanced colorimetry biosensor based on a toehold-containing three-way junction (TWJ) induced multiple isothermal exponential amplification and the CRISPR/Cas14a (C-TEC) biosensor. The TWJ template was designed as a Y–X–Y structure. In the presence of the target, the formation of toehold-containing TWJ complex induced primer extension, leading to the generation of amplified single-stranded DNA; this amplified DNA could then bind to either the free TWJ template for EXPAR reaction or the toehold of the TWJ complex for toehold-mediated strand displacement, thereby enabling the recycling of the target. The amplification products could trigger CRISPR/Cas14a for efficient trans-cleavage and release the magnetically bound gold nanoparticle probes for colorimetry detection. Using Mycobacterium tuberculosis 16S rDNA as the target, the proposed C-TEC could detect 16S rDNA down to 50 fM by the naked eye and 20.71 fM by UV–vis detector at 520 nm within 90 min under optimal conditions. We successfully applied this biosensor to clinical isolates of Mycobacterium tuberculosis. In addition, the C-TEC biosensor also showed feasibility for the detection of RNA viruses. In conclusion, the proposed C-TEC is a convenient, fast, and versatile platform for visual detection of pathogen DNA/RNA and has potential clinical applications.
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