title: Microfluidic LIPS for serum antibody detection: demonstration of a rapid test for HSV-2 infection.
abstract: There is great interest in point-of-care antibody testing for the diagnosis of infectious and autoimmune diseases. As a first step in the development of self-contained and miniaturized devices for highly quantitative antibody detection, we demonstrate the application of Luciferase Immunoprecipitation Systems (LIPS) technology in a microfluidic format. Protein A/G was immobilized on the walls of PDMS-glass microchannels of 500 nL volume. The assay proceeds with the simultaneous introduction of plasma and Renilla luciferase-tagged antigens. Following washing, coelenterazine substrate was added and bound antigen-luciferase measured by chemiluminescence. Total assay time, including rinsing and detection, is under 10 min. Using these stable microfluidic devices, high diagnostic performance (100% sensitivity and 100% specificity) was achieved for the diagnosis of HSV-2 infection. Based on these findings, the LIPS microfluidic format should readily lend itself to automation and the transfer to portable instrumentation.
authors.affiliation: Microfabrication and Microfluidics Unit, Biomedical Engineering and Physical Science Shared Resource, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bldg. 13, Rm. 3N18, 13 South Drive, Bethesda, MD 20892-5766, USA.
authors.forename: Adnan
authors.initials: A
authors.lastname: Zubair
authors.forename: Peter D
authors.initials: PD
authors.lastname: Burbelo
authors.forename: Ludovic G
authors.initials: LG
authors.lastname: Vincent
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Iadarola
authors.forename: Paul D
authors.initials: PD
authors.lastname: Smith
authors.forename: Nicole Y
authors.initials: NY
authors.lastname: Morgan
mesh_terms: D000914:Antibodies, Viral / Q000097:blood* / Q000302:isolation & purification*; D004867:Equipment Design; D006558:Herpes Genitalis / Q000175:diagnosis*; D018258:Herpesvirus 2, Human / Q000302:isolation & purification; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D008156:Luciferases / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000914:Antibodies, Viral; D008156:Luciferases