Shrestha Kiran, Kim Seongryeong, Han Jiyeon, Zhang Meng, Parajuli Sajjan, Cho Gyoujin
Department of Biophysics, Institute of Quantum Biology, Sungkyunkwan University, Suwon, 16419, South Korea.
Department of Intelligent Precision Healthcare Convergence, Sungkyunkwan University, Suwon, 16419, South Korea.
Microsyst Nanoeng. 2025 Jul 15;11(1):143. doi: 10.1038/s41378-025-00972-w.
Photothermal conversion-based quantitative polymerase chain reaction (qPCR) is a fast, sensitive, and accurate method to diagnose infectious diseases. However, they have bottlenecks in test throughput scalability, cumbersome oil cover, and a lack of multi-target capability. Here, the authors present an infectious disease diagnostic device with rapid photothermal conversion-based efficient reverse transcription (RT)-qPCR assays on a multi-target chip (idream-qPCR). The authors innovate an off-axis mirror-based three-channel fluorescence intensity measurement method, enabling concurrent non-contact temperature control of 16 mini-well reaction chambers for qPCRs without the necessity of actuating parts. A transparent adhesive film on a graphite mixed polydimethylsiloxane (PDMS)-based PCR chip with mini-wells avoids contamination and bubbles to achieve 16 RT-qPCRs (40 photothermal cycles) within 17 min. Finally, idream-qPCR is validated by amplifying severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) N1 72 bp, RdRP 100 bp, and E 113 bp genes using Fluorescein amidites (FAM), Carboxytetramethylrhodamine (TAMRA), and Cyanine5 (CY5) fluorescent dyes, respectively, with 102.5% efficiency and a limit-of-detection (LoD) equivalent to 0.85 copies/µL. idream-qPCR can be efficiently used to prevent the spread of infectious diseases.
基于光热转换的定量聚合酶链反应（qPCR）是一种快速、灵敏且准确的传染病诊断方法。然而，它们在测试通量可扩展性、繁琐的油覆盖以及缺乏多靶点检测能力方面存在瓶颈。在此，作者展示了一种传染病诊断设备，该设备在多靶点芯片上基于光热转换进行高效逆转录（RT）-qPCR检测（idream-qPCR）。作者创新了一种基于离轴镜的三通道荧光强度测量方法，能够对16个用于qPCR的微孔反应室进行并行非接触温度控制，而无需驱动部件。基于石墨混合聚二甲基硅氧烷（PDMS）的带有微孔的PCR芯片上的透明胶膜可避免污染和气泡，从而在17分钟内实现16次RT-qPCR（40个光热循环）。最后，通过分别使用荧光素酰胺（FAM）、羧基四甲基罗丹明（TAMRA）和花青素5（CY5）荧光染料扩增严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的N1 72bp、RdRP 100bp和E 113bp基因，对idream-qPCR进行了验证，效率为102.5%，检测限（LoD）相当于0.85拷贝/微升。idream-qPCR可有效用于预防传染病的传播。
