一个基于单RPA-CRISPR/Cas12a驱动器G4-hemin自组的裸眼生物传感系统用于Mycobacterium tuberculosis
Ting Yuan1,2, Jianbo Yuan1,2, Jian Huang1,2
1Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi, China.
Frontiers in chemistry
|August 25, 2025
概括
一个新的生物传感系统可以快速,灵敏地检测出Mycobacterium tuberculosis (MTB). 这种临床试验结合了RPA和CRISPR/Cas12a,可在不到一个小时的时间内对MTB核酸进行视觉识别.
科学领域:
- 生物技术
- 分子诊断
- 生物传感
背景情况:
- 准确的结核菌 (MTB) 鉴定对于结核病 (TB) 控制至关重要.
- 传统的MTB诊断面临的局限性包括低灵敏度,高成本和复杂的程序.
- 需要简单,快速和灵敏的MTB诊断方法.
研究的目的:
- 开发一台生物传感系统,以快速和灵敏地检测MTB.
- 克服现有的MTB检测诊断方法的局限性.
- 为MTB核酸测试创建一个无设备的视觉检测方法.
主要方法:
- 开发了一种整合复合酶聚合酶放大 (RPA) 和CRISPR/Cas12a技术的单系统.
- 用甘油作为分相屏障来优化RPA和CRISPR/ Cas12a的相互作用.
- 通过CRISPR/Cas12a介导的ssDNA探针的裂变形成G4-胺纳米酶,使色度检测成为可能.
主要成果:
- 生物传感系统实现了MTB的超敏感肉眼检测,检测极限为10个副本/μL.
- 在不需要复杂仪器的情况下,测试在60分钟内提供了结果.
- 用104个患者样本进行的临床验证显示与标准诊断方法具有很高的一致性.
结论:
- 开发的生物传感系统提供了一种简单,快速和高度灵敏的MTB检测方法.
- 这种方法结合了RPA,CRISPR/Cas12a和G4-heminDNAzyme以实现有效的视觉识别.
- 该系统显示了在资源有限的环境中进行MTB核酸检测的巨大潜力.
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