Department of Biotechnology, University of Bahri, Khartoum, Sudan
Department of Biotechnology, University of Bahri, Khartoum, Sudan.
J Clin Microbiol. 2021 Feb 18;59(3). doi: 10.1128/JCM.00745-20.
Infectious diseases are one of the most intimidating threats to human race, responsible for an immense burden of disabilities and deaths. Rapid diagnosis and treatment of infectious diseases offers a better understanding of their pathogenesis. According to the World Health Organization, the ideal approach for detecting foreign pathogens should be rapid, specific, sensitive, instrument-free, and cost-effective. Nucleic acid pathogen detection methods, typically PCR, have numerous limitations, such as highly sophisticated equipment requirements, reagents, and trained personnel relying on well-established laboratories, besides being time-consuming. Thus, there is a crucial need to develop novel nucleic acid detection tools that are rapid, specific, sensitive, and cost-effective, particularly ones that can be used for versatile point-of-care diagnostic applications. Two new methods exploit unpredicted properties of CRISPR-Cas effectors, turning activated nucleases into basic amplifiers of a specific nucleic acid binding event. These effectors can be attached to a diversity of reporters and utilized in tandem with isothermal amplification approaches to create sensitive identification in multiple deployable field formats. Although still in their beginning, SHERLOCK and DETECTR technologies are potential methods for rapid detection and identification of infectious diseases, with ultrasensitive tests that do not require complicated processing. This review describes SHERLOCK and DETECTR technologies and assesses their properties, functions, and prospective to become the ultimate diagnostic tools for diagnosing infectious diseases and curbing disease outbreaks.
传染病是人类最可怕的威胁之一,造成了巨大的残疾和死亡负担。快速诊断和治疗传染病可以更好地了解其发病机制。根据世界卫生组织的说法,检测外来病原体的理想方法应该是快速、特异、敏感、无仪器且具有成本效益。核酸病原体检测方法,通常是 PCR,存在许多限制,例如对高度复杂的设备、试剂和依赖于成熟实验室的训练有素的人员的要求,此外还需要耗费时间。因此,迫切需要开发新型的核酸检测工具,这些工具应具有快速、特异、敏感和具有成本效益的特点,特别是那些可以用于多功能即时诊断应用的工具。两种新方法利用了 CRISPR-Cas 效应物的意外特性,将激活的核酸酶转变为特定核酸结合事件的基本放大器。这些效应物可以与各种报告分子结合使用,并与等温扩增方法结合使用,以在多种可部署的现场格式中进行敏感识别。尽管仍处于起步阶段,但 SHERLOCK 和 DETECTR 技术是用于快速检测和识别传染病的潜在方法,具有无需复杂处理的超灵敏测试。本文描述了 SHERLOCK 和 DETECTR 技术,并评估了它们的特性、功能以及成为诊断传染病和控制疾病爆发的最终诊断工具的潜力。