Next-generation CRISPR-based diagnostic tools for human diseases
Next-generation CRISPR-based diagnostic tools for human diseases
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DOI：
10.1016/j.trac.2023.117328
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发表时间：
2023-09
期刊：
影响因子：
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通讯作者：
Ting Wang;Ziwei Wang;Linlin Bai;Xingcai Zhang;Hongyu Zhang;Jia Feng;Cheng Qian;Yongming Wang-Yongmi
中科院分区：
文献类型：
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作者：
Ting Wang;Ziwei Wang;Linlin Bai;Xingcai Zhang;Hongyu Zhang;Jia Feng;Cheng Qian;Yongming Wang-Yongmi
人类疾病的准确诊断对于有效预防、临床治疗和预后至关重要。世界卫生组织（WHO）已经提供了分子诊断方法的指导方针，建议理想的方法应符合ASSURED标准：然而，传统的分子检测方法具有一定的局限性，包括需要昂贵的试剂和设备，熟练的技术人员，冗长的处理时间，劳动强度大，灵敏度低，在偏远地区和实时场景中的适用性有限。快速发展的CRISPR/Cas系统通过提供低成本，多功能和强大的工具箱，彻底改变了分子诊断。在这篇综述中，我们将首先介绍CRISPR工具箱在诊断中的一般工作原理。然后，我们将介绍基于CRISPR/Cas系统的各种核酸和非核酸检测方法，用于诊断感染性和非感染性疾病，特别关注单核苷酸多态性（SNP）的检测。最后，我们讨论了CRISPR分子诊断所面临的挑战，并期待通过合理的设计和多学科工程在疾病诊断中获得更多突破性的应用，以实现多重、免预扩增、即时检测（POCT）和疾病的连续监测。
Accurate diagnosis of human diseases is crucial for effective prevention, clinical treatment, and prognosis. The World Health Organization (WHO) has provided guidelines for molecular diagnostic methods, suggesting that the ideal method should meet the ASSURED criteria: affordable, sensitive, specific, user-friendly, rapid, robust, equipment-free, and deliverable to end-users.However, traditional molecular detection methods have certain limitations, including the need for expensive reagents and equipment, skilled technicians, lengthy processing time, high labor intensity, low sensitivity, and limited applicability in remote areas and real-time scenarios. The rapidly evolving CRISPR/Cas systems have revolutionized molecular diagnostics by providing a low-cost, versatile, and powerful toolbox. In this review, we will first introduce the general working principle of the CRISPR toolbox in diagnosis. We will then present various nucleic acid and non-nucleic acid detection methods based on CRISPR/Cas systems for diagnosing both infectious and non-infectious diseases, with a particular focus on the detection of single nucleotide polymorphisms (SNPs). Finally, we discuss the challenges faced by CRISPR molecular diagnostics and expect more groundbreaking applications in disease diagnostics through rational design and multidisciplinary engineering to enable multiplexed, preamplification-free, point-of-care testing (POCT) and continuous monitoring of disease.