病原核酸CRISPR-Cas可视化检测方法研究进展

Advances in visual detection of pathogen nucleic acids by CRISPR-Cas

摘要:

可视化检测是通过肉眼判读检测结果，不依赖复杂的光学检测系统的一种检测方法，大多基于可见光、气压变化和迁移距离等肉眼可直接观察的信号快速获取结果，被广泛应用于体外诊断行业。成簇规律间隔短回文重复序列及相关蛋白(clustered regularly interspaced short palindromic repeats and CRISPR-associated protein, CRISPR-Cas)系统以其反应条件温和，无需热循环等控制，且具备较强的信号放大能力，在发展即时检测(point of care testing, POCT)技术中具有非常大的潜力。可视化检测与CRISPR-Cas系统的联用，极大地降低了核酸检测对实验室基础设施、精密仪器及专业人员的依赖程度，推动了核酸POCT技术和方法的发展。本文综述了CRISPR-Cas系统核酸可视化检测技术的信号输出方式及特点，并对该可视化检测技术应用中存在的问题进行了讨论，最后对其未来的临床转化进行了展望，为研发CRISPR-Cas可视化检测方法提供了参考。

Abstract:

Visual detection is a technique for evaluating the results through visual judgment without relying on complex optical detection systems. It obtains results quickly based on signals, such as visible light, changes in air pressure, and migration distance, that can be directly observed by naked eyes, being widely used in the in vitro diagnostics industry. The CRISPR-Cas system has the potential to be used in the development of point of care testing (POCT) technologies due to the advantages of mild reaction conditions, no need for thermal cycling or other control measures, and a robust signal amplification capability. In recent years, the combination of visual detection and CRISPR-Cas has significantly reduced the need for laboratory infrastructures, precision instruments, and specialized personnel for nucleic acid detection. This has promoted the development of POCT technology and methods for nucleic acids. This article summarizes the signal output modes and characteristics of the visual detection of nucleic acid by CRISPR-Cas and discusses the issues in the application. Finally, its future clinical translation is envisioned with a view to informing the development of CRISPR-Cas visualization assays.