Liu Ming, Wang Lei, Zhou Jiaxi, Miao Guijun, Zhang Lulu, Gao Lizeng, Qiu Xianbo
Institute of Microfluidic Chip Development in Biomedical Engineering, School of Information Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.
CAS Engineering Laboratory for Nanozyme, Institute of Biophysics, Chinese Academic of Science, Beijing, 100101, China.
Analyst. 2025 Jul 7;150(14):3158-3169. doi: 10.1039/d5an00237k.
Influenza viruses spread rapidly, seriously threatening human health, for example, causing serious and even deadly respiratory diseases, and weakening the global economy. Influenza virus detection methods include molecular detection and immunoassay, and compared to molecular detection, the traditional lateral flow immunoassay is widely used in point-of-care testing (POCT) due to its advantages of fast detection, low cost and ease of use. However, these assays are often limited by trivial operational procedure, specialized skills, and long detection time. To address these challenges and enable rapid and highly sensitive pathogen detection, we developed a centrifugal analyzing platform and a centrifugal microfluidic chip integrated with a nanozyme lateral flow strip for automated detection of H1N1 viral antigens. The signal amplification using nanozyme-based enzyme-linked immunosorbent assay (ELISA) significantly improves the sensitivity and shortens the assay time. A peak searching algorithm was utilized to semi-quantitatively analyze the detection and control lines on the lateral flow strip of the nanozyme, which showed a 2.5-fold increase in the lower limit of detection compared to the eye-observation-based assay. In the automated detection system, the centrifugal microfluidic chip sequentially delivers the sample and color development solution to the lateral flow strip, facilitating fully automated operation and minimizing the risk of manual errors. Experimental results show that the manual operation takes only 1 minute, and the total testing time is only 16 minutes. This study highlights a multifunctional platform for automated nanozyme-based ELISA assay, offering broad applicability in medical diagnostics, environmental monitoring, and food safety testing.
流感病毒传播迅速，严重威胁人类健康，例如会引发严重甚至致命的呼吸道疾病，并削弱全球经济。流感病毒检测方法包括分子检测和免疫测定，与分子检测相比，传统的侧向流动免疫测定因其检测速度快、成本低且使用方便等优点，在即时检测（POCT）中被广泛应用。然而，这些检测方法常常受到操作流程繁琐、需要专业技能以及检测时间长的限制。为应对这些挑战并实现快速且高灵敏度的病原体检测，我们开发了一种离心分析平台以及一种集成了纳米酶侧向流动试纸条的离心微流控芯片，用于自动检测H1N1病毒抗原。使用基于纳米酶的酶联免疫吸附测定（ELISA）进行信号放大，显著提高了灵敏度并缩短了检测时间。利用一种峰值搜索算法对纳米酶侧向流动试纸条上的检测线和控制线进行半定量分析，结果显示与基于肉眼观察的检测方法相比，检测下限提高了2.5倍。在自动检测系统中，离心微流控芯片将样品和显色液依次输送到侧向流动试纸条上，便于实现全自动化操作并将人为误差风险降至最低。实验结果表明，手动操作仅需1分钟，总检测时间仅为16分钟。本研究突出了一个基于纳米酶的多功能自动ELISA检测平台，在医学诊断、环境监测和食品安全检测方面具有广泛的适用性。
