Department of Applied Chemistry, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan.
Analyst. 2024 Sep 23;149(19):4932-4939. doi: 10.1039/d4an00905c.
Although CRISPR-based nucleic acid detection has great potential in point-of-care testing due to its simplicity, it has been rarely integrated into paper-based analytical devices (PADs), which are attractive platforms to simplify assays. This work introduces a CRISPR-assisted nucleic acid quantification approach integrated into a PAD with signal readout by a personal glucose meter (PGM). Retention of magnetic beads by filter paper and pre-deposition of all required reagents by freeze-drying stabilized with trehalose enabled the indirect quantification of human papilloma virus (HPV) DNA through a PGM readout without complicated user intervention and complex reagent handling. The calculated limit of detection was 57 pM, which is comparable with other amplification-free CRISPR-based assays detecting nucleic acids. The fully integrated device exhibited good storage stability for up to 4 weeks, suggesting its applicability toward practical point-of-care nucleic acid quantification.
尽管基于 CRISPR 的核酸检测因其简单性而在即时检测中有很大的潜力，但它很少被整合到基于纸张的分析设备 (PAD) 中，而 PAD 是简化检测的有吸引力的平台。本工作介绍了一种将 CRISPR 辅助的核酸定量方法集成到 PAD 中的方法，该方法通过个人血糖仪 (PGM) 进行信号读出。滤纸截留磁性珠和用海藻糖稳定的冻干预沉积所有必需的试剂，使得能够通过 PGM 读出进行人乳头瘤病毒 (HPV) DNA 的间接定量，而无需复杂的用户干预和复杂的试剂处理。计算出的检测限为 57 pM，与其他无扩增的基于 CRISPR 的检测核酸的方法相当。完全集成的设备表现出长达 4 周的良好储存稳定性，表明其适用于实际的即时检测核酸定量。
