Conventional colorimetric enzyme-linked immunosorbent assay (ELISA) is a time-consuming laboratory assay that is not very sensitive and consumes a large amount of samples. Herein, the development of a reusable, cost-effective, and eco-friendly poly(methyl methacrylate) (PMMA)/paper hybrid plug-and-play (PnP) device for high-sensitivity immunoassay by analyte enrichment and efficient passing-through washing has been reported. The PMMA device has multiple slots where a pre-patterned paper substrate can be inserted. The sample flows back-and-forth through a low-cost, 3D paper substrate within the PMMA channels, thereby enhancing the amount of analyte adsorbed and dramatically increasing the sensitivity while decreasing the assay time. After the enrichment assay, the paper substrate can simply be pulled out of the device, and the results can be qualitatively viewed with the naked eye or scanned through a simple desktop scanner for quantitative analysis. The paper substrate can be replaced with a new substrate so that the device can be reused. The limits of detection (LODs) of 200 pg/mL for immunoglobulin G (IgG) and 270 pg/mL for hepatitis B surface antigen (HBsAg) were obtained. This IgG assay is at least 10 times more sensitive than commercial ELISA kits. In addition, the PnP ELISA exhibited a significant increase in the linear dynamic range from 3 orders of magnitude in a common paper-based device to a wide range of six orders of magnitude in the PnP hybrid device. This reusable PnP device has great potential for the low-cost yet high-sensitivity detection of infectious diseases, cancers, and other important biomolecules.

传统的比色酶联免疫吸附试验 (ELISA) 是一种耗时的实验室检测方法，灵敏度不高，需要大量样品。在此，开发了一种可重复使用、经济高效且环保的聚（甲基丙烯酸甲酯）（PMMA）/纸混合即插即用（PnP）设备，用于通过分析物富集和高效通过洗涤进行高灵敏度免疫测定已经被报告了。PMMA 设备有多个插槽，可以插入预先图案化的纸基材。样品在 PMMA 通道内的低成本 3D 纸基材上来回流动，从而增加了分析物的吸附量并显着提高了灵敏度，同时缩短了分析时间。富集测定后，纸基材可以简单地从设备中拉出，并且结果可以用肉眼定性查看或通过简单的桌面扫描仪进行扫描以进行定量分析。纸质基材可以更换为新的基材，以便设备可以重复使用。免疫球蛋白 G (IgG) 的检测限 (LOD) 为 200 pg/mL，乙型肝炎表面抗原 (HBsAg) 的检测限 (LOD) 为 270 pg/mL。这种 IgG 检测的灵敏度至少是商业 ELISA 试剂盒的 10 倍。此外，PnP ELISA 的线性动态范围显着增加，从普通纸质设备的 3 个数量级到 PnP 混合设备的 6 个数量级。这种可重复使用的 PnP 设备在传染病、癌症和其他重要生物分子的低成本、高灵敏度检测方面具有巨大潜力。