Paper-based analytical device for point-of-care nucleic acid quantification combining CRISPR/Cas12a and a personal glucose meter,Analyst

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Paper-based analytical device for point-of-care nucleic acid quantification combining CRISPR/Cas12a and a personal glucose meter
Analyst ( IF 3.6 ) Pub Date : 2024-07-29 , DOI: 10.1039/d4an00905c
Yohei Tanifuji ₁ , Guodong Tong ₁ , Yuki Hiruta ₁ , Daniel Citterio ₁

Affiliation

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.

中文翻译：

结合 CRISPR/Cas12a 和个人血糖仪的用于即时核酸定量的纸质分析装置

尽管基于 CRISPR 的核酸检测由于其简单性而在即时检测中具有巨大潜力，但它很少被集成到纸质分析设备 (PAD) 中，而纸质分析设备是简化检测的有吸引力的平台。这项工作介绍了一种集成到 PAD 中的 CRISPR 辅助核酸定量方法，并通过个人血糖仪 (PGM) 读取信号。通过滤纸保留磁珠，并通过用海藻糖稳定的冷冻干燥来预沉积所有所需试剂，可以通过 PGM 读数间接定量人乳头状瘤病毒 (HPV) DNA，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，与其他基于 CRISPR 的无扩增核酸检测方法相当。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护理点核酸定量。

更新日期：2024-07-29

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