In this study, a microfluidic-based system utilizing colorimetric loop-mediated isothermal amplification (LAMP) is introduced for the quantitative analysis of nucleic acid targets. This system offers a user-friendly and cost-effective platform for the multiplexed genetic diagnosis of various infectious diseases across multiple samples. It includes time-lapse imaging equipment for capturing images of the microfluidic device during the LAMP assay and a hue-based quantitative analysis software to analyze the LAMP reaction, streamlining diagnostic procedures. An electric pipette was used to simplify the loading of samples and LAMP reagents into the device, allowing easy operation even by untrained individuals. The hue-based analysis software employs efficient image processing and post-processing techniques to calculate DNA amplification curves based on color changes in multiple reaction chambers. This software automates several tasks, such as identifying reaction chamber areas from time-lapse images, quantifying color information within each chamber, correcting baselines of DNA amplification curves, fitting experimental data to theoretical curves, and determining the threshold time for each curve. To validate the developed system, conventional off-chip LAMP assays were conducted with a 25 μL reaction mixture in 0.2 mL polymerase chain reaction (PCR) tubes using a real-time turbidimeter. The results indicated that the threshold time obtained using the colorimetric LAMP assay in the developed system is comparable to that obtained with real-time turbidity measurements in PCR tubes, demonstrating the system's capability for quantitative analysis of target nucleic acids, including those from human herpesviruses.

中文翻译：

---

基于微流体的定量分析系统，使用比色环介导的等温扩增对人类病毒感染进行多重基因诊断

在这项研究中，引入了一种利用比色环介导等温扩增（LAMP）的微流体系统来对核酸靶标进行定量分析。该系统为跨多个样本的各种传染病的多重基因诊断提供了一个用户友好且具有成本效益的平台。它包括用于在 LAMP 测定过程中捕获微流体装置图像的延时成像设备和用于分析 LAMP 反应的基于色调的定量分析软件，从而简化诊断程序。使用电动移液器简化了将样品和 LAMP 试剂加载到设备中的过程，即使未经培训的人员也可以轻松操作。基于色调的分析软件采用高效的图像处理和后处理技术，根据多个反应室中的颜色变化计算 DNA 扩增曲线。该软件可自动执行多项任务，例如从延时图像中识别反应室区域、量化每个反应室内的颜色信息、校正 DNA 扩增曲线的基线、将实验数据拟合到理论曲线以及确定每条曲线的阈值时间。为了验证所开发的系统，使用实时浊度计在 0.2 mL 聚合酶链反应 (PCR) 管中使用 25 μL 反应混合物进行传统的片外 LAMP 测定。结果表明，在开发的系统中使用比色 LAMP 测定获得的阈值时间与在 PCR 管中实时浊度测量获得的阈值时间相当，证明了该系统对目标核酸（包括来自人类疱疹病毒的核酸）进行定量分析的能力。