Zhang Shanshan, Duan Mingyuan, Li Shuang, Hou Jie, Qin Ting, Teng Zhanwei, Hu Jianhe, Zhang Huihui, Xia Xiaojing
College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, China.
College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, China.
Diagn Microbiol Infect Dis. 2023 Oct 1;108(1):116097. doi: 10.1016/j.diagmicrobio.2023.116097.
Rapid detection of pathogenic microorganisms is key to the epidemiologic identification, prevention and control of disease in the field of public health. PCR-based pathogen detection methods have been widely used because they overcome the time-consuming issues encountered in traditional culture-based methods, including the limited detecting window-phase of immunological detection. However, the requirement for precise temperature-controlled thermal cyclers severely limits the application of these methods in resource-limited areas. Recombinase polymerase amplification (RPA) is a new type of nucleic acid amplification technology that can amplify DNA or RNA at a constant temperature. It has the advantages of simple operation, high specificity and sensitivity and a short detection time. In recent years, a number of alternative methods for pathogenic microorganism detection have been developed by combining microfluidic technology with RPA. Through the design of chip structures, optimization of injection modes, and utilization of multiple detection and quantification methods, the integration of pathogen nucleic acid extraction, amplification and detection is achieved, and this approach is suitable for the rapid detection of pathogenic microorganisms in various environments. In this review, we compare different nucleic acid amplification techniques, explain the principle of RPA technology, detection methods, and applications for pathogen microorganism detection and describe future direction of RPA application. These methods increase the ability to rapidly screen pathogenic microorganisms, thus improving the management of infectious diseases in the field of public health.
快速检测致病微生物是公共卫生领域疾病流行病学识别、预防和控制的关键。基于聚合酶链反应（PCR）的病原体检测方法已被广泛应用，因为它们克服了传统基于培养的方法中遇到的耗时问题，包括免疫检测的检测窗口期有限。然而，对精确温度控制的热循环仪的要求严重限制了这些方法在资源有限地区的应用。重组酶聚合酶扩增（RPA）是一种新型的核酸扩增技术，可在恒温下扩增DNA或RNA。它具有操作简单、特异性和灵敏度高以及检测时间短的优点。近年来，通过将微流控技术与RPA相结合，开发了许多用于致病微生物检测的替代方法。通过芯片结构设计、进样模式优化以及多种检测和定量方法的利用，实现了病原体核酸提取、扩增和检测的一体化，该方法适用于各种环境中致病微生物的快速检测。在这篇综述中，我们比较了不同的核酸扩增技术，解释了RPA技术的原理、检测方法及其在病原体微生物检测中的应用，并描述了RPA应用的未来方向。这些方法提高了快速筛查致病微生物的能力，从而改善了公共卫生领域传染病的管理。
