Kim MinGin, Yun Sung-Hun, Kim Sun-Hee, Kim Jong-Dae
Thermo Fisher Scientific, South San Francisco, CA 94080, USA.
School of Software, Hallym University, Chuncheon-si 24252, Republic of Korea.
Sensors (Basel). 2025 May 17;25(10):3159. doi: 10.3390/s25103159.
We present a compact and cost-effective real-time PCR system designed for point-of-care testing (POCT), utilizing a PCB-based disposable chip and an open-platform CMOS camera. The system integrates precise thermal cycling with software-synchronized fluorescence detection and provides real-time analysis through a dedicated user interface. To minimize cost and complexity, a polycarbonate reaction chamber was integrated with a PCB-based heater and thermistor. A slanted LED illumination setup and an open-platform USB camera were employed for fluorescence imaging. Signal alignment was enhanced using device-specific region-of-interest (ROI) tracking based on copper pad corner detection. Thermal cycling performance achieved a heating rate of 8.0 °C/s and a cooling rate of -9.3 °C/s, with steady-state accuracy within ±0.1 °C. Fluorescence images exhibited high dynamic range without saturation, and the 3σ-based ROI correction method improved signal reliability. System performance was validated using DNA standard (10 copies), yielding consistent amplification curves with a Ct standard deviation below 0.3 cycles. These results demonstrate that the proposed system enables rapid, accurate, and reproducible nucleic acid detection, making it a strong candidate for field-deployable molecular diagnostics.
我们展示了一种紧凑且经济高效的实时聚合酶链反应(PCR)系统,该系统专为即时检测(POCT)设计,采用基于印刷电路板(PCB)的一次性芯片和开放平台的互补金属氧化物半导体(CMOS)相机。该系统将精确的热循环与软件同步荧光检测相结合,并通过专用用户界面提供实时分析。为了将成本和复杂性降至最低,聚碳酸酯反应室与基于PCB的加热器和热敏电阻集成在一起。采用倾斜的发光二极管(LED)照明设置和开放平台的通用串行总线(USB)相机进行荧光成像。基于铜焊盘角点检测,使用特定设备的感兴趣区域(ROI)跟踪增强了信号对齐。热循环性能实现了8.0℃/秒的加热速率和-9.3℃/秒的冷却速率,稳态精度在±0.1℃以内。荧光图像呈现出高动态范围且无饱和现象,基于3σ的ROI校正方法提高了信号可靠性。使用DNA标准品(10个拷贝)对系统性能进行了验证,得到了一致的扩增曲线,其循环阈值(Ct)标准偏差低于0.3个循环。这些结果表明,所提出的系统能够实现快速、准确和可重复的核酸检测,使其成为现场可部署分子诊断的有力候选者。