Zhang Tianyi, Wang Xiaoqin, Jiang Yingtao, Zhang Yunyun, Zhao Shuhao, Hu Jian, Hu Fei, Peng Niancai
State Key Laboratory for Manufacturing Systems Engineering, School of Instrument Science and Technology, Xi'an Jiaotong University, Xi'an, 710054, China.
Clinical Laboratory, First Affiliated Hospital of Xi 'an Jiaotong University, Xi'an, 710061, China.
Anal Chim Acta. 2025 Feb 8;1338:343593. doi: 10.1016/j.aca.2024.343593. Epub 2024 Dec 27.
Nucleic acid testing is the most effective detection method currently available for the diagnosis of respiratory infectious diseases. However, the conventional real-time fluorescent quantitative PCR technique, which is regarded as the gold standard method for nucleic acid detection, presents significant challenges for implementation in home self-testing and popularization in underdeveloped regions due to its rigorous experimental standards. It is therefore clear that an easy-to-use, miniaturized nucleic acid testing technology and products for nonprofessionals are of great necessity to define the pathogens and assist in controlling disease transmission. (87) RESULTS: In this study, we propose a strategy for self-testing of respiratory pathogen nucleic acid that is oriented towards the public and user-friendly. The proposed system integrates the processes of extraction-free nucleic acid release, RPA isothermal amplification, and CRISPR fluorescence detection into a compact configuration. A microfluidic testing chip actuated by air pouches and a battery/USB-powered reusable device has been developed to enable simultaneous detection of internal reference genes and viral targets in a fully enclosed condition. The system allows for sample-in, and result-out testing in less than 30 min with a detection limit of 2 copies/μL. Additionally, a straightforward signal-light-based result display method has been developed to make it easy and intuitive for users to access the results. Furthermore, freeze-drying reagent is introduced to guarantee the storage and transportation of testing chips in ambient conditions. (135) SIGNIFICANCE: This work presents a miniaturized, portable, and highly sensitive nucleic acid detection system, where simple operating procedures have been designed for unskilled users. It is our belief that the testing system developed in this work is well suited for home-based self-testing and infection diagnosis in resource-limited areas, due to the above-mentioned advantages. (52).
核酸检测是目前诊断呼吸道传染病最有效的检测方法。然而，传统的实时荧光定量PCR技术虽被视为核酸检测的金标准方法，但因其严格的实验标准，在家庭自测及欠发达地区推广方面面临重大挑战。因此，显然非常有必要开发一种便于非专业人员使用的小型化核酸检测技术及产品，以明确病原体并助力控制疾病传播。（87）结果：在本研究中，我们提出了一种面向公众且用户友好的呼吸道病原体核酸自测策略。所提出的系统将免提取核酸释放、RPA等温扩增及CRISPR荧光检测过程整合为紧凑结构。已开发出一种由气囊驱动的微流控检测芯片及一个由电池/USB供电的可重复使用装置，以实现在完全封闭条件下同时检测内参基因和病毒靶点。该系统可在不到30分钟内实现进样及出结果检测，检测限为2拷贝/μL。此外，还开发了一种基于信号灯的直观结果显示方法，方便用户获取结果。此外，引入冻干试剂以确保检测芯片在环境条件下的储存和运输。（135）意义：本研究成果展示了一种小型化、便携式且高灵敏度的核酸检测系统，为非专业用户设计了简单的操作程序。我们认为，鉴于上述优势，本研究开发的检测系统非常适合在资源有限地区进行家庭自测及感染诊断。（52）
