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CRISPR-Cas系统在病原微生物检测中的应用
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Abstract:
快速、灵敏和特异性地检测病原微生物，在临床诊断和传染病控制中具有重要意义。早期准确检测是快速控制疫区疫情的有效措施，尤其是在缺乏有效治疗和疫苗的情况下。聚合酶链式反应(PCR)作为常用的核酸检测技术和疾病诊断的“金标准”，有着高灵敏度的优势，但同时也存在以牺牲特异性为代价的现象。酶联免疫吸附试验(ELISA)是一种快速、特异性强的蛋白质和小分子诊断工具。然而，灵敏度低和样品预处理复杂的操作步骤极大地限制了该方法现场检测的应用。因此，快速、灵敏和特异性的检测技术成为了急需解决的要点，随着技术的应用和发展，基于CRISPR-Cas的生物传感系统的优异性能在开发病原微生物诊断技术方面引起了人们的关注。本文综述了CRISPR/Cas系统在病原微生物检测的作用机制及原理，总结了新型检测技术的优缺点并对应用发展前景进行展望。
Rapid, sensitive, and specific detection of pathogenic microorganisms is crucial for clinical diagnosis and infectious disease control. Early and accurate detection is an effective measure to quickly control epidemic outbreaks, especially in the absence of effective treatments and vaccines. Polymerase Chain Reaction (PCR) is a commonly used nucleic acid testing technology and the “gold standard” for disease diagnosis with high sensitivity. However, it often sacrifices specificity. Enzyme-Linked Immunosorbent Assay (ELISA) is a rapid and highly specific diagnostic tool for proteins and small molecules. Nevertheless, its low sensitivity and complex sample preprocessing steps greatly limit its application in on-site testing. Therefore, the development of rapid, sensitive, and specific detection technologies has become an urgent need. With the application and advancement of technology, the excellent performance of CRISPR-Cas based biosensing systems has attracted attention in the development of pathogenic microorganism diagnostic techniques. This article reviews the mechanisms and principles of CRISPR/Cas systems in pathogenic microorganism detection, summarizes the advantages and disadvantages of novel detection technologies, and provides an outlook on their future applications.
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