A paper/polymer hybrid microfluidic microplate for rapid quantitative detection of multiple disease biomarkers.
A paper/polymer hybrid microfluidic microplate for rapid quantitative detection of multiple disease biomarkers.
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DOI：
10.1038/srep30474
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发表时间：
2016-07-26
影响因子：
4.6
通讯作者：
Li X
中科院分区：
文献类型：
--
作者：
Sanjay ST;Dou M;Sun J;Li X
酶联免疫吸附试验（ELISA）是目前应用最广泛的实验室疾病诊断方法之一。然而，在资源匮乏的环境中进行ELISA受到孵育时间长、大量珍贵试剂和设备齐全的实验室的限制。在此，我们开发了一种简单的，小型化的纸/PMMA（聚（甲基丙烯酸甲酯））混合微流控微板，用于低成本，高通量和即时（POC）的传染病诊断。多孔纸在流通微孔中的新用途有助于快速抗体/抗原固定和有效洗涤，避免复杂的表面修饰。顶部的试剂输送通道可以简单地将试剂转移到多个微孔中，从而避免重复的手动移液和昂贵的机器人。用肉眼可在1小时内观察到比色ELISA的结果。通过计算办公室扫描仪扫描图像的亮度来实现定量分析。对人血清中免疫球蛋白G（IgG）和B型肝炎表面抗原（HBsAg）进行了定量分析，结果可靠。在不使用任何专用设备的情况下，IgG和HBsAg的检测限分别为1.6 ng/mL和1.3 ng/mL，与使用专用设备的商业ELISA试剂盒相当。我们设想这种简单的POC混合微孔板可以在各种生物测定中具有广泛的应用，特别是在资源有限的环境中。
Enzyme linked immunosorbent assay (ELISA) is one of the most widely used laboratory disease diagnosis methods. However, performing ELISA in low-resource settings is limited by long incubation time, large volumes of precious reagents, and well-equipped laboratories. Herein, we developed a simple, miniaturized paper/PMMA (poly(methyl methacrylate)) hybrid microfluidic microplate for low-cost, high throughput, and point-of-care (POC) infectious disease diagnosis. The novel use of porous paper in flow-through microwells facilitates rapid antibody/antigen immobilization and efficient washing, avoiding complicated surface modifications. The top reagent delivery channels can simply transfer reagents to multiple microwells thus avoiding repeated manual pipetting and costly robots. Results of colorimetric ELISA can be observed within an hour by the naked eye. Quantitative analysis was achieved by calculating the brightness of images scanned by an office scanner. Immunoglobulin G (IgG) and Hepatitis B surface Antigen (HBsAg) were quantitatively analyzed with good reliability in human serum samples. Without using any specialized equipment, the limits of detection of 1.6 ng/mL for IgG and 1.3 ng/mL for HBsAg were achieved, which were comparable to commercial ELISA kits using specialized equipment. We envisage that this simple POC hybrid microplate can have broad applications in various bioassays, especially in resource-limited settings.