克里斯普尔双酶分裂触发了SARS-CoV-2双基因检测的DNA和RNA基质分裂
Tong Jiang1,2, Runde Liu1,2, Jilu Shen1,2
1The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China.
Journal of medical virology
|September 11, 2023
概括
这项研究引入了一个快速的视觉诊断平台,用于使用CRISPR-Cas12a/Cas13a和RT-RPA同时检测SARS-CoV-2ORF1ab和N基因. 这种新的方法在不到20分钟的时间内实现了高灵敏度和精度.
科学领域:
- 分子生物学分子生物学
- 生物技术是生物技术.
- 传染病诊断 传染病诊断 传染病诊断
背景情况:
- 冠状病毒2019 (COVID-19) 的广泛传播需要快速和准确的诊断方法来缓解传播.
- 目前的诊断方法可能面临速度,灵敏度或同时检测多个病毒点的局限性.
研究的目的:
- 开发和验证一个多重诊断平台,用于快速,超敏感,视觉和同时检测严重急性呼吸综合征冠状病毒2 (SARS-CoV-2) ORF1ab和N基因.
- 整合聚类定期间隔短平行体重复 (CRISPR) -Cas12a/Cas13a双酶系统与多重反转录酶-重组酶聚合酶放大 (RT-RPA) 进行增强的诊断能力.
主要方法:
- 开发了一种结合CRISPR-Cas12a/Cas13a双酶消化与多重RT-RPA的新型诊断平台.
- 该系统利用Cas12a和Cas13a酶,分别对SARS-CoV-2 N和ORF1ab基因进行特定的识别和分离.
- 使用光信号或CRISPR双消化试验条进行了切割产品的检测,从而实现了视觉识别.
主要成果:
- 多重诊断平台在不到20分钟的时间内完成了快速分析.
- 该方法实现了高达200个副本/毫升的超敏感检测极限.
- 在105个临床鼻拭片样本中的验证显示,与定量逆转录-聚合酶链反应相比,100%的积极和95.7%的负预测值协议.
结论:
- 开发的CRISPR-Cas12a/Cas13a-RT-RPA平台提供了一种新的方法,用于快速,视觉和同时检测SARS-CoV-2.
- 该方法提供了一个灵敏而准确的诊断工具,有可能在传染病监测中得到广泛应用.
- 该平台的效率和高性能为SARS-CoV-2的临床诊断提供了显著的进步.
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