College of Public Health, Jilin Medical University, Jilin, 132013, People's Republic of China.
Medical College, Yanbian University, Yanji, 136200, People's Republic of China.
Int J Nanomedicine. 2023 Nov 6;18:6311-6331. doi: 10.2147/IJN.S434197. eCollection 2023.
Recent, outbreaks of new emergency zoonotic diseases have prompted an urgent need to develop fast, accurate, and portable screening assays for pathogen infections. Recombinase polymerase amplification (RPA) is sensitive and specific and can be conducted at a constant low temperature with a short response time, making it especially suitable for on-site screening and making it a powerful tool for preventing or controlling the spread of zoonoses. This review summarizes the design principles of RPA-based biosensors as well as various signal output or readout technologies involved in fluorescence detection, lateral flow assays, enzymatic catalytic reactions, spectroscopic techniques, electrochemical techniques, chemiluminescence, nanopore sequencing technologies, microfluidic digital RPA, and clustered regularly interspaced short palindromic repeats/CRISPR-associated systems. The current status and prospects of the application of RPA-based biosensors in zoonoses screening are highlighted. RPA-based biosensors demonstrate the advantages of rapid response, easy-to-read result output, and easy implementation for on-site detection, enabling development toward greater portability, automation, and miniaturization. Although there are still problems such as high cost with unstable signal output, RPA-based biosensors are increasingly becoming one of the most important means of on-site pathogen screening in complex samples involving environmental, water, food, animal, and human samples for controlling the spread of zoonotic diseases.
最近，新出现的紧急人畜共患传染病的爆发，促使人们迫切需要开发用于病原体感染的快速、准确和便携式筛选检测方法。重组酶聚合酶扩增（RPA）具有灵敏度高和特异性强的特点，并且可以在恒定的低温下进行，具有较短的响应时间，因此特别适合现场筛选，是预防或控制人畜共患病传播的有力工具。本综述总结了基于 RPA 的生物传感器的设计原理，以及涉及荧光检测、侧向流动分析、酶催化反应、光谱技术、电化学技术、化学发光、纳米孔测序技术、微流控数字 RPA 和簇状规则间隔短回文重复序列/CRISPR 相关系统的各种信号输出或读出技术。突出了基于 RPA 的生物传感器在人畜共患病筛选中的应用的现状和前景。基于 RPA 的生物传感器具有快速响应、易于读取结果输出和易于实现现场检测的优势，朝着更大的便携性、自动化和小型化方向发展。尽管存在信号输出不稳定、成本高等问题，但基于 RPA 的生物传感器越来越成为现场病原体筛选的重要手段之一，用于控制人畜共患病在涉及环境、水、食品、动物和人体样本等复杂样本中的传播。
