Zeng Wu, Li Bao, Chen Yanjing, Lin Baobao, Gu Xuanyu, Liu Peng, Zhang Yan
School of Biomedical Engineering, Tsinghua University, Beijing 100084, China.
Changping Laboratory, Beijing 102206, China.
Anal Chem. 2025 Sep 2;97(34):18469-18478. doi: 10.1021/acs.analchem.5c01644. Epub 2025 Aug 18.
The cocirculation and infection of SARS-CoV-2 and influenza A viruses significantly impact the transmission dynamics of infectious diseases and increase the public health burden on communities. Nucleic acid amplification testing (NAAT), the gold standard for pathogen detection, has been widely used in point-of-care testing (POCT). However, consumer-grade NAAT-based medical POCT products tailored to resource-limited settings remain scarce. In this study, we developed a finger-actuated visual microfluidic chip with a colorimetric detection system based on reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) to enable efficient and cost-effective detection of SARS-CoV-2 and influenza A. During on-site testing, 280-340 μL of oropharyngeal swab sample is first added to the microfluidic chip through a dropper. The sample is then evenly distributed across four separate reaction chambers within the chip by the force driven by finger pressure. Through quantitative and qualitative experiments, we verified the feasibility and versatility of the chip and demonstrated that this system exhibits high sensitivity, with a limit of detection (LoD) of 100 copies/μL in extraction-free devices, while effectively preventing cross-contamination between individual reaction chambers. Testing with 20 mock clinical swab samples showed that the system achieved 100% detection rate and specificity. Thus, we believe the finger-actuated microfluidic system, with its low cost, ease of use, and versatile detection capabilities, can offer a powerful POCT tool for timely viral infection confirmation during the influenza season.
严重急性呼吸综合征冠状病毒2(SARS-CoV-2)和甲型流感病毒的共同传播和感染对传染病的传播动态有重大影响,并增加了社区的公共卫生负担。核酸扩增检测(NAAT)作为病原体检测的金标准,已广泛应用于即时检测(POCT)。然而,针对资源有限环境量身定制的基于消费级NAAT的医疗POCT产品仍然稀缺。在本研究中,我们开发了一种基于逆转录环介导等温扩增(RT-LAMP)比色检测系统的手指驱动视觉微流控芯片,以实现对SARS-CoV-2和甲型流感的高效且经济高效的检测。在现场检测期间,首先通过滴管将280 - 340 μL的口咽拭子样本添加到微流控芯片中。然后,在手指压力驱动的力作用下,样本在芯片内的四个独立反应腔中均匀分布。通过定量和定性实验,我们验证了该芯片的可行性和通用性,并证明该系统具有高灵敏度,在无提取装置中的检测限(LoD)为100拷贝/μL,同时有效防止各个反应腔之间的交叉污染。对20个模拟临床拭子样本的检测表明,该系统的检测率和特异性均达到100%。因此,我们相信这种手指驱动的微流控系统,因其低成本、易用性和通用的检测能力,可为流感季节及时确认病毒感染提供强大的POCT工具。