title: A paper/polymer hybrid microfluidic microplate for rapid quantitative detection of multiple disease biomarkers.
abstract: 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.
authors.affiliation: Department of Chemistry, University of Texas at El Paso, 500 West University Ave, El Paso, Texas, 79968, USA.
authors.forename: Sharma T
authors.initials: ST
authors.lastname: Sanjay
authors.affiliation: Department of Chemistry, University of Texas at El Paso, 500 West University Ave, El Paso, Texas, 79968, USA.
authors.forename: Maowei
authors.initials: M
authors.lastname: Dou
authors.affiliation: Border Biomedical Research Center, University of Texas at El Paso, 500 West University Ave, El Paso, Texas, 79968, USA.
authors.forename: Jianjun
authors.initials: J
authors.lastname: Sun
authors.affiliation: Department of Chemistry, University of Texas at El Paso, 500 West University Ave, El Paso, Texas, 79968, USA.|Border Biomedical Research Center, University of Texas at El Paso, 500 West University Ave, El Paso, Texas, 79968, USA.|Biomedical Engineering, University of Texas at El Paso, 500 West University Ave, El Paso, Texas, 79968, USA.
authors.forename: XiuJun
authors.initials: X
authors.lastname: Li
mesh_terms: D015415:Biomarkers / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D006514:Hepatitis B Surface Antigens / Q000097:blood; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007211:Indoles / Q000737:chemistry / Q000378:metabolism; D044085:Microfluidics / Q000379:methods*; D009580:Nitroblue Tetrazolium / Q000737:chemistry / Q000378:metabolism; D010209:Paper; D011108:Polymers / Q000737:chemistry*; D019904:Polymethyl Methacrylate / Q000737:chemistry; D013997:Time Factors
chemical_list: D015415:Biomarkers; D006514:Hepatitis B Surface Antigens; D007074:Immunoglobulin G; D007211:Indoles; D011108:Polymers; D009580:Nitroblue Tetrazolium; C035455:5-bromo-4-chloro-3-indoxyl phosphate; D019904:Polymethyl Methacrylate