Department of Instrument Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Engineering Research Center for Intelligent Diagnosis and Treatment Instrument, Key Laboratory of Thin Film and Microfabrication (Ministry of Education), Shanghai Jiao Tong University, Shanghai, 200240, China.
School of Naval Architecture, Ocean & Civil Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
Anal Bioanal Chem. 2020 Apr;412(11):2517-2528. doi: 10.1007/s00216-020-02475-1. Epub 2020 Feb 18.
Paper-based immunoassays are effective methods that employ microfluidic paper-based analytical devices (μPADs) for the rapid, simple, and accurate quantification of analytes in point-of-care diagnosis. In this study, we developed a wax-printed multilayered μPAD for the colorimetric detection of carcinoembryonic antigen (CEA), where the device contained a movable and rotatable detection layer to allow the μPAD to switch the state of the sample solutions, i.e., flowing or storing in the sensing zones. A smartphone with a custom-developed program served as an automated colorimetric reader to capture and analyze images from the μPAD, before calculating and displaying the test results. After optimizing the crucial conditions for the assay, the proposed method exhibited a wide linear dynamic range from 0.5 to 70 ng/mL, with a low CEA detection limit of 0.015 ng/mL. The clinical performance of this method was successfully validated using 50 positive and 40 negative human serum samples, thereby demonstrating the high sensitivity of 98.0% and specificity of 97.5% in the detection of CEA. The proposed method is greatly simplified compared with the cumbersome steps required for traditional immunoassays, but without any loss of accuracy and stability, as well as reducing the time needed to detect CEA. Complex and bulky instruments are replaced with a smartphone. The proposed detection platform could potentially be applied in point-of-care testing. Graphical abstract.
基于纸的免疫分析是一种有效的方法，它采用微流控纸基分析器件 (μPAD) 快速、简单、准确地定量分析即时诊断中的分析物。在本研究中，我们开发了一种蜡印多层 μPAD 用于癌胚抗原 (CEA) 的比色检测，其中该装置包含一个可移动和可旋转的检测层，以允许 μPAD 切换样品溶液的状态，即流动或存储在感应区。智能手机与自定义开发的程序一起用作自动比色读取器，用于捕获和分析 μPAD 的图像，然后计算并显示测试结果。在优化了该检测方法的关键条件后，该方法表现出从 0.5 到 70ng/mL 的宽线性动态范围，CEA 的检测限低至 0.015ng/mL。使用 50 份阳性和 40 份阴性人血清样本成功验证了该方法的临床性能，从而证明了检测 CEA 的高灵敏度为 98.0%和特异性为 97.5%。与传统免疫分析所需的繁琐步骤相比，该方法大大简化，但准确性和稳定性没有任何损失，同时还缩短了检测 CEA 的时间。复杂且庞大的仪器被智能手机取代。所提出的检测平台有可能应用于即时检测。
