Digital CRISPR Systems for the Next Generation of Nucleic Acid Quantification.
Digital CRISPR Systems for the Next Generation of Nucleic Acid Quantification.
复制标题
DOI：
10.1016/j.trac.2023.116917
复制
发表时间：
2023-01
期刊：
影响因子：
--
通讯作者：
Anthony J Politza;Reza Nouri;W. Guan
中科院分区：
文献类型：
--
作者：
Anthony J Politza;Reza Nouri;W. Guan
数字CRISPR （dCRISPR）检测是一种新兴的分子诊断平台。数字平台引入了绝对定量，提高了批量CRISPR分析的灵敏度。dCRISPR系统具有超特异性靶向、等温操作和快速检测等特点，为引领分子诊断领域做好了充分准备。这里我们总结了CRISPR检测中常用的Cas蛋白。研究了数字检测方法和关键性能因素。我们形成了三种策略来构建dCRISPR系统的景观：(1)无扩增，(2)分区内扩增，(3)两阶段扩增。我们还通过检测极限（LOD）、测试时间和性能值（FOM）比较了所有系统的性能。本工作总结了数字CRISPR平台的细节，以指导未来的发展。我们预计，LOD和动态范围的改进将使dCRISPR成为下一代分子生物传感的领先平台。
Digital CRISPR (dCRISPR) assays are an emerging platform of molecular diagnostics. Digital platforms introduce absolute quantification and increased sensitivity to bulk CRISPR assays. With ultra-specific targeting, isothermal operation, and rapid detection, dCRISPR systems are well-prepared to lead the field of molecular diagnostics. Here we summarized the common Cas proteins used in CRISPR detection assays. The methods of digital detection and critical performance factors are examined. We formed three strategies to frame the landscape of dCRISPR systems: (1) amplification free, (2) in-partition amplification, and (3) two-stage amplification. We also compared the performance of all systems through the limit of detection (LOD), testing time, and figure of merit (FOM). This work summarizes the details of digital CRISPR platforms to guide future development. We envision that improvements to LOD and dynamic range will position dCRISPR as the leading platform for the next generation of molecular biosensing.