通讯作者: 姚亚超, yaoyc@gd2h.org.cn

DOI：10.12201/bmr.202410.00045

Advances in the application of amplification-free CRISPRCas cascade signal amplification technology

• 摘要：快速准确的分子诊断在精准医疗、环境监测、食品安全等生命科学领域发挥重要作用。基于成簇规则间隔短回文重复序列（clustered regularly interspaced short palindromic repeats，CRISPR）及相关蛋白（CRISPR-associated protein，Cas）组成的CRISPR及相关酶（CRISPR/Cas）系统因其具有的高灵敏度、高特异性、便捷性及快速性等优势，已成为一种非常有效的分子诊断工具。然而，基于CRISPR/Cas系统的检测要通过预扩增步骤提高靶标浓度，这可能会面临非特异性扩增、气溶胶污染、检测周期长和操作复杂等问题。因此人们开发出了各种免扩增检测策略，但这些免扩增检测策略因其灵敏度低而不能满足大多数疾病的诊断需求，而一系列的研究表明级联信号放大策略在提高基于CRISPR/cas的检测方法的灵敏度方面具有广阔的应用前景。本文综述了免扩增CRISPR/Cas级联信号放大技术的主要策略和应用进展。

  关键词： CRISPR/Cas; 免扩增检测; 超灵敏检测; 信号放大

   

  Abstract: Rapid and accurate molecular diagnosis plays an important role in life science fields such as precision medicine, environmental monitoring, and food safety. CRISPR/Cas system based on clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas) has become a very effective molecular diagnostic tool due to its advantages of high sensitivity, high specificity, convenience and rapidity. CRISPR/Cas system has become a very effective molecular diagnostic tool due to its advantages of high sensitivity, high specificity, convenience and speed. However, CRISPR/Cas system-based assays increase the target concentration through a pre-amplification step, which may also generate problems such as non-specific amplification, aerosol contamination, complex operation and long assay cycle. Therefore, various amplification-free detection strategies have been developed, but the low sensitivity of these amplification-free detection strategies cannot meet the diagnostic needs of most diseases. A series of studies have demonstrated that the cascade signal amplification strategy has a broad application prospect in improving the sensitivity of CRISPR/cas-based assays, and the amplification-free CRISPR/Cas cascade signal amplification technology in disease diagnosis is reviewed. The application of amplification-free CRISPR/Cas cascade signal amplification technology in disease diagnosis is reviewed, and the main strategies of amplification-free CRISPR/Cas cascade signal amplification technology are highlighted.

  Key words: CRISPR/Cas; amplification-free detection; ultrasensitive detection; signal amplification

  提交时间：2024-10-16

  版权声明：作者本人独立拥有该论文的版权，预印本系统仅拥有论文的永久保存权利。任何人未经允许不得重复使用。
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• 序号	提交日期	编号	操作
  1	2024-09-12	bmr.202410.00045V1	下载

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