Zhao Xue, Gao Jiale, Gu Yijing, Teng Zheng, Zhang Xi, Wu Huanyu, Chen Xin, Chen Min, Kong Jilie
Virus Testing Laboratory, Pathogen Testing Center, Shanghai Municipal Center for Disease Control and Prevention, Shanghai 201100, China.
School of Life Science and Technology, Tongji University, Shanghai 200092, China.
Biosensors (Basel). 2025 Aug 4;15(8):506. doi: 10.3390/bios15080506.
: Influenza viruses are major pathogens responsible for respiratory infections and pose significant risks to densely populated urban areas. RT-qPCR has made substantial contributions in controlling virus transmission during previous COVID-19 epidemics, but it faces challenges in terms of detection time for large sample sizes and susceptibility to nucleic acid contamination. : Our study designed loop-mediated isothermal amplification primers for three common influenza viruses: A/H3N2, A/H1N1, and B/Victoria, and utilized a 4-channel microfluidic chip to achieve simultaneous detection. The chip initiates amplification by centrifugation and allows testing of up to eight samples at a time. : By creating a closed amplification system in the microfluidic chip, aerosol-induced nucleic acid contamination can be prevented through physically isolating the reaction from the operating environment. The chip can specifically detect A/H1N1, A/H3N2, and B/Victoria and has no signal for other common respiratory viruses. The testing process can be completed within 1 h and can be sensitive to viral RNA at concentrations as low as 10 ng/μL for A/H1N1 and A/H3N2 and 10 ng/μL for B/Victori. A total of 296 virus swab samples were further analyzed using the microfluidic chip method and compared with the classical qPCR method, which resulted in high consistency. : Our chip enables faster detection of influenza virus and avoids nucleic acid contamination, which is beneficial for POCT establishment and has lower requirements for the operating environment.
流感病毒是引起呼吸道感染的主要病原体，对人口密集的城市地区构成重大风险。逆转录定量聚合酶链反应（RT-qPCR）在先前新冠疫情期间对控制病毒传播做出了重大贡献，但在大样本量检测时间和对核酸污染的敏感性方面面临挑战。我们的研究针对三种常见流感病毒：甲型H3N2、甲型H1N1和乙型Victoria设计了环介导等温扩增引物，并利用四通道微流控芯片实现同步检测。该芯片通过离心启动扩增，一次可检测多达八个样本。通过在微流控芯片中创建封闭的扩增系统，通过将反应与操作环境物理隔离，可以防止气溶胶引起的核酸污染。该芯片可以特异性检测甲型H1N1、甲型H3N2和乙型Victoria，对其他常见呼吸道病毒无信号。检测过程可在1小时内完成，对甲型H1N1和甲型H3N2病毒RNA浓度低至10 ng/μL以及乙型Victoria病毒RNA浓度低至10 ng/μL均敏感。使用微流控芯片方法对总共296份病毒拭子样本进行了进一步分析，并与经典qPCR方法进行比较，结果具有高度一致性。我们的芯片能够更快地检测流感病毒并避免核酸污染，这有利于即时检测（POCT）的建立，并且对操作环境要求较低。
