title: Highly Sensitive Two-Dimensional Paper Network Incorporating Biotin-Streptavidin for the Detection of Malaria.
abstract: Recently, two-dimensional paper networks have been developed to enable multistep assays to be performed in a lateral flow format. These devices have been used to perform simple enzyme linked immunoassays on paper. However, these devices have yet to incorporate more complex immunoassays, including the use of streptavidin-biotin detection strategies. Here we present a modified two-dimensional paper network capable of consecutively delivering six reagents. The device requires only a single user step and delivers (i) the sample, (ii) the biotinylated detection antibody, (iii) streptavidin horseradish peroxidase, (iv) a wash buffer, (v) a colorimetric substrate, and (vi) a final wash buffer. To demonstrate the utility of this approach we designed an assay to detect the malaria protein Pf HRP2. Using this platform, we were able to achieve a limit-of-detection equivalent to that of a traditional 96-well plate sandwich ELISA. In addition to improvements in the limit-of-detection, the inclusion of streptavidin-biotin simplifies the development of similar tests for other targets.
authors.affiliation: Department of Bioengineering, Rice University , MS-142, 6100 Main Street, Houston, Texas 77005, United States.
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Grant
authors.affiliation: Department of Bioengineering, Rice University , MS-142, 6100 Main Street, Houston, Texas 77005, United States.
authors.forename: Chelsey A
authors.initials: CA
authors.lastname: Smith
authors.affiliation: Department of Bioengineering, Rice University , MS-142, 6100 Main Street, Houston, Texas 77005, United States.|University of Virginia School of Medicine , Charlottesville, Virginia 22908, United States.
authors.forename: Kristine
authors.initials: K
authors.lastname: Karvonen
authors.affiliation: Department of Bioengineering, Rice University , MS-142, 6100 Main Street, Houston, Texas 77005, United States.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Richards-Kortum
mesh_terms: D000953:Antigens, Protozoan / Q000032:analysis* / Q000737:chemistry; D001710:Biotin / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D006735:Horseradish Peroxidase / Q000737:chemistry; D007202:Indicators and Reagents; D057230:Limit of Detection; D008288:Malaria / Q000175:diagnosis; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods; D015800:Protozoan Proteins / Q000032:analysis* / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry*
chemical_list: D000953:Antigens, Protozoan; C514457:HRP-2 antigen, Plasmodium falciparum; D007202:Indicators and Reagents; D015800:Protozoan Proteins; D001710:Biotin; D019809:Streptavidin; D006735:Horseradish Peroxidase
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