Jing Wenwen, Zhang Tong, Min Xiangyang, Li Xin, Jin Kai, Feng Meng, Sui Guodong, Luo Liulin, Cheng Xunjia
Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P. R. China.
Department of Clinical Laboratory, Shanghai. East Hospital, School of Medicine, Tong Ji University, Shanghai 200120, P. R. China.
ACS Omega. 2024 Sep 6;9(37):38989-38997. doi: 10.1021/acsomega.4c05489. eCollection 2024 Sep 17.
The outbreak poses health risks to community residents. However, it still has limitations for current clinical diagnostic methods (qPCR nucleic acid assay or IgM immunoassay), including specialized handling, expensive equipment, prolonged turnaround time, and false positives and negatives, highlighting the need to improve clinical diagnostic methods. Herein, we present a novel centrifugal microfluidics-based method for rapidly diagnosing infections (CHAMP system). This user-friendly method combines CRISPR/Cas12b and real-time loop-mediated isothermal amplification (LAMP) in a one-pot reaction, offering high sensitivity, specificity, and simplicity for methodology. By adding fully automated nucleic acid magnetic bead-extracted samples to a prepackaged centrifugal microfluidics chip, 48 samples can be automated tested simultaneously within 15 to 60 min at 60 °C. 427 clinical nasopharyngeal swab specimens were used for validation, demonstrating good positive and negative predictive values and good diagnostic sensitivity, specificity, and significant time savings. This method is particularly suitable for detecting low nucleic acid copies of samples.
此次疫情对社区居民构成健康风险。然而，目前的临床诊断方法（定量聚合酶链反应核酸检测或免疫球蛋白M免疫测定）仍存在局限性，包括需要专门处理、设备昂贵、周转时间长以及存在假阳性和假阴性结果，这凸显了改进临床诊断方法的必要性。在此，我们提出了一种基于离心微流控技术的新型感染快速诊断方法（CHAMP系统）。这种用户友好型方法将CRISPR/Cas12b和实时环介导等温扩增技术（LAMP）结合在一个反应体系中，具有高灵敏度、特异性且方法简单。通过将全自动核酸磁珠提取的样本添加到预包装的离心微流控芯片中，可在60℃下15至60分钟内同时对48个样本进行自动检测。使用427份临床鼻咽拭子标本进行验证，结果显示该方法具有良好的阳性和阴性预测值，以及良好的诊断灵敏度、特异性，且显著节省时间。该方法特别适用于检测样本中低核酸拷贝数的情况。
