Naorungroj Sarida, Mahardika Ignasia Handipta, Huh Eunjin, Yukird Jutiporn, Pasomsub Ekawat, Chailapakul Orawon, Shin Kwanwoo
Electrochemistry and Optical Spectroscopy Center of Excellence (EOSCE), Department of Chemistry, Faculty of Science, Chulalongkorn University, Phayathai Road, Pathumwan, Bangkok, 10330, Thailand.
Department of Chemistry and Institute of Biological Interfaces, Sogang University, Seoul, 04107, Republic of Korea.
Mikrochim Acta. 2025 May 28;192(6):381. doi: 10.1007/s00604-025-07228-4.
A fluorescence origami paper-based analytical device (Flu-oPAD) based on a strand displacement assay as a point-of-care testing (POCT) sensing platform for DNA detection is presented. This device facilitates multiple steps in a single device, including sample loading, incubation, and washing. The detection zone was immobilized with a 6-FAM-modified probe (F-probe), which formed a complex with a BHQ1-modified probe (Q-probe) to minimize background signals. In the presence of target DNA, hybridization with the F-probe releases the Q-probe, resulting in a fluorescence response proportional to the target DNA concentration. The sensor exhibits good selectivity for target DNA, with a linearity range from 0.1 nM to 10 μM and an experimental detection limit of 0.1 nM, completing all procedures within 15 min. The real-world applicability is demonstrated by successfully detecting complementary DNA (cDNA) from the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a cause of the COVID-19 pandemic. This study further introduces a multi-array Flu-oPAD for simultaneous detection of SARS-CoV-2 ORF1ab, N, and E gene cDNAs. This multi-array Flu-oPAD is applied to nasopharyngeal swab samples, showing good agreement with the standard RT-PCR method. Overall, the developed Flu-oPAD has shown great potential as an effective POCT tool for DNA screening. It offers simplicity, portability, accessibility, and cost-effectiveness, offering its potential impact on addressing pressing healthcare needs.
本文介绍了一种基于链置换分析的荧光折纸纸质分析装置（Flu-oPAD），作为一种用于DNA检测的即时检测（POCT）传感平台。该装置在单个设备中实现了多个步骤，包括样品加载、孵育和洗涤。检测区固定有6-羧基荧光素修饰的探针（F-探针），它与BHQ1修饰的探针（Q-探针）形成复合物以最小化背景信号。在存在目标DNA的情况下，与F-探针杂交会释放Q-探针，从而产生与目标DNA浓度成比例的荧光响应。该传感器对目标DNA具有良好的选择性，线性范围为0.1 nM至10 μM，实验检测限为0.1 nM，可在15分钟内完成所有程序。通过成功检测严重急性呼吸综合征冠状病毒2（SARS-CoV-2，即COVID-19大流行的病原体）的互补DNA（cDNA），证明了其在实际应用中的适用性。本研究还引入了一种多阵列Flu-oPAD，用于同时检测SARS-CoV-2的ORF1ab、N和E基因cDNA。这种多阵列Flu-oPAD应用于鼻咽拭子样本，与标准RT-PCR方法显示出良好的一致性。总体而言，所开发的Flu-oPAD作为一种有效的DNA筛查POCT工具显示出巨大潜力。它具有简单、便携、易获取和成本效益高的特点，对满足紧迫的医疗需求具有潜在影响。
