基于RPA-CRISPR/Cas12a-SPM和深度学习的DNA病毒检测系统
Changyue Liu1, Zhengyang Lei1, Lijin Lian2
1Center of Precision Medicine and Healthcare, Tsinghua-Berkeley Shenzhen Institute; Institute of Biopharmaceutics and Health Engineering, Tsinghua Shenzhen International Graduate School.
Journal of visualized experiments : JoVE
|May 27, 2024
概括
这项研究介绍了一种使用复合酶聚合酶放大 (RPA) 和CRISPR/Cas12a的快速,临床点的DNA病毒检测系统. 该系统能够在40分钟内检测到像青病毒3 (FV3) 这样的病毒.
科学领域:
- 分子生物学分子生物学
- 生物技术是生物技术.
- 病毒学 病毒学
背景情况:
- DNA 病毒对公众健康构成重大威胁.
- 快速和灵敏的检测方法对于及时诊断和控制至关重要.
- 现有的方法往往需要专门的设备和训练有素的人员,限制了点的护理应用.
研究的目的:
- 开发一种快速,灵敏和特定序列的DNA病毒检测系统,用于医疗点 (POC) 使用.
- 将重组酶聚合酶放大 (RPA) 与CRISPR/Cas12a系统集成,以提高检测能力.
- 将智能手机显微镜和人工智能 (AI) 纳入自动化分析和分类.
主要方法:
- 利用复合酶聚合酶放大 (RPA) 进行向DNA放大.
- 使用CRISPR/Cas12a系统进行序列特异性识别和DNA记者附带裂解.
- 集成的便携式智能手机显微镜 (SPM) 用于光成像和深度学习模型,用于人工智能辅助的分类.
主要成果:
- 实现了DNA病毒的微量检测,青病毒3 (FV3) 的检测极限 (LoD) 低至10aM.
- 整个检测过程,包括放大和分析,在40分钟内完成.
- 在使用人工智能辅助分析对阳性和阴性样本的二进制分类中证明了高准确性.
- 该系统在不需要熟练操作人员或大型实验室仪器的情况下被证明是有效的.
结论:
- 开发的RPA-CRISPR/Cas12a-SPM系统提供了一个便携式,灵敏和快速的解决方案,用于在护理点检测DNA病毒.
- 整合人工智能可以提高诊断准确性和可访问性.
- 这项技术在防止DNA病毒在各种环境中传播方面具有显著的潜力.
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