Chen Yingwei, Zhao Ran, Hu Xiaobo, Wang Xueliang
Department of Quality Control Material R&D, Shanghai Center for Clinical Laboratory, Shanghai, PR China; Department of Molecular Diagnostic Innovation Technology, Shanghai Academy of Experimental Medicine, Shanghai, PR China.
Department of Quality Control Material R&D, Shanghai Center for Clinical Laboratory, Shanghai, PR China; Department of Molecular Biology, Shanghai Center for Clinical Laboratory, Shanghai, PR China.
Anal Chim Acta. 2025 Jan 22;1336:343295. doi: 10.1016/j.aca.2024.343295. Epub 2024 Oct 1.
The current pandemic of 2022 global mpox (formerly known as monkeypox), caused by infection with monkeypox virus (MPXV), has now reached over 120 countries. This constitutes a critical public health issue requiring effective disease management and surveillance. Rapid and reliable diagnosis is conducive to the control of infection, early intervention, and timely treatment. Clinical laboratories use various conventional diagnostic methods for detecting MPXV, including PCR, which can be regarded as a gold-standard diagnostic method. However, the application of PCR is limited by its requirements for high-cost equipment, skilled professionals, and a laboratory setting.
Clustered regularly interspaced short palindromic repeats (CRISPR)-based diagnostic systems have provided promising prospects for the rapid, sensitive, and specific detection of infectious diseases, especially in point-of-care settings. Over the past 2 years, an increasing number of researchers have concentrated on the application of the CRISPR method to mpox diagnosis. In the majority of cases, a two-step method was chosen, with CRISPR/Cas12a and recombinase polymerase amplification (RPA) as pre-amplification methods, followed by a fluorescence readout. Different strategies have been applied to overcome the encountered limitations of CRISPR detection, but no consensus on an integrated solution has been achieved. Thus, the application of the CRISPR/Cas system in mpox detection requires further exploration and improvement.
This review discusses contemporary studies on MPXV CRISPR detection systems and the strategies proposed to address the challenges faced by CRISPR diagnosis with the hope of helping the development of CRISPR detection methods and improving pathogen detection technologies.
由猴痘病毒（MPXV）感染引起的2022年全球猴痘（原称猴天花）大流行现已蔓延至120多个国家。这构成了一个关键的公共卫生问题，需要有效的疾病管理和监测。快速可靠的诊断有助于控制感染、早期干预和及时治疗。临床实验室使用各种传统诊断方法检测MPXV，包括PCR，它可被视为金标准诊断方法。然而，PCR的应用受到其对高成本设备、技术熟练的专业人员和实验室环境要求的限制。
基于成簇规律间隔短回文重复序列（CRISPR）的诊断系统为传染病的快速、灵敏和特异性检测提供了广阔前景，尤其是在即时检测环境中。在过去两年中，越来越多的研究人员专注于将CRISPR方法应用于猴痘诊断。在大多数情况下，选择了两步法，以CRISPR/Cas12a和重组酶聚合酶扩增（RPA）作为预扩增方法，随后进行荧光读数。已应用不同策略来克服CRISPR检测中遇到的局限性，但尚未就综合解决方案达成共识。因此，CRISPR/Cas系统在猴痘检测中的应用需要进一步探索和改进。
本综述讨论了关于MPXV的CRISPR检测系统的当代研究以及为应对CRISPR诊断面临的挑战而提出的策略，希望有助于CRISPR检测方法的发展并改进病原体检测技术。
