Sanjay Sharma T, Li Meihan, Zhou Wan, Li Xiaochun, Li XiuJun
Department of Chemistry and Biochemistry, University of Texas at El Paso, 500 West University Ave, El Paso, TX 79968 USA.
College of Biomedical Engineering, Taiyuan University of Technology, 030024 Taiyuan, Shanxi China.
Microsyst Nanoeng. 2020 Jun 15;6:28. doi: 10.1038/s41378-020-0143-5. eCollection 2020.
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测定法（ELISA）是一种耗时的实验室检测方法，灵敏度不高且消耗大量样本。在此，报道了一种可重复使用、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）装置，用于通过分析物富集和高效直通洗涤进行高灵敏度免疫测定。PMMA装置有多个插槽，可插入预图案化的纸质基底。样品在PMMA通道内通过低成本的三维纸质基底来回流动，从而增加吸附的分析物量，显著提高灵敏度，同时缩短检测时间。富集测定后，只需将纸质基底从装置中取出，结果可用肉眼定性查看，或通过简单的桌面扫描仪扫描进行定量分析。纸质基底可更换为新的基底，以便装置能够重复使用。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）的检测限为270 pg/mL。这种IgG检测方法的灵敏度至少比商业ELISA试剂盒高10倍。此外，PnP ELISA的线性动态范围从普通纸质装置的3个数量级显著增加到PnP混合装置的6个数量级的宽范围。这种可重复使用的PnP装置在低成本高灵敏度检测传染病、癌症和其他重要生物分子方面具有巨大潜力。
