title: Development of a novel microfluidic immunoassay for the detection of Helicobacter pylori infection.
abstract: The miniaturization of laboratory processes offers substantial advantages over traditional techniques in terms of cost, speed, and potential for multistage automation. To date, only a few studies have reported successful microfluidics-based immunoassays, most of which rely on fluorescence detection technologies. The goal of this study was to develop a poly(dimethylsiloxane) microfluidics-based immunoassay methodology and a versatile colorimetric quantification scheme for the detection of visual colour changes resulting from immune reactions in microchannels. The novel immunoassay technique was applied towards the detection of Helicobacter pylori infection using 20 human serum samples of known infection status, and results compared with conventional nitrocellulose membrane-based dot-ELISA. The microchannel immunoassay reliably detected H. pylori antigens in quantities on the order of 10 ng, which provides a sensitivity of detection comparable to conventional dot-blot assays. Sensitivity was 100%, specificity was 90%, positive predictive value 91%, and negative predictive value 100%, with an overall accuracy of 95%. The software developed generated results that were consistent with visual observations and by automatically taking into account background intensity changes, the software minimized subjectivity. Volumes of solutions used were 100-fold less compared with conventional immunoassays. Miniaturization of the ELISA using this technique provides a means for the accurate diagnosis of microbial infection while minimizing waste production.
authors.affiliation: Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.
authors.forename: Frank Y H
authors.initials: FY
authors.lastname: Lin
authors.forename: Mahdi
authors.initials: M
authors.lastname: Sabri
authors.forename: David
authors.initials: D
authors.lastname: Erickson
authors.forename: Javad
authors.initials: J
authors.lastname: Alirezaie
authors.forename: Dongqing
authors.initials: D
authors.lastname: Li
authors.forename: Philip M
authors.initials: PM
authors.lastname: Sherman
mesh_terms: D000942:Antigens, Bacterial / Q000097:blood*; D003124:Colorimetry / Q000379:methods; D016481:Helicobacter Infections / Q000175:diagnosis*; D016480:Helicobacter pylori / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D008832:Microchemistry / Q000379:methods
chemical_list: D000942:Antigens, Bacterial