title: A thin layer-based amperometric enzyme immunoassay for the rapid and sensitive diagnosis of respiratory syncytial virus infections.
abstract: A simple electrochemical sandwich immunoassay involving a polystyrene microarray slide coated with monoclonal capture antibodies and carbon screen-printed sensors (SPS) was designed for the rapid diagnosis of respiratory syncytial virus (RSV). The detection of the antibody-antigen complex formation relied on the use of a horseradish peroxidase conjugate. Its chronoamperometric measurement detection was performed by confining a droplet of H(2)O(2)/3,3',5,5'-tetramethylbenzidine enzyme substrate/mediator solution within a thin layer between one spot of the microarray and the surface of one screen-printed electrochemical cell. The accumulation of the enzyme product in the thin film of liquid enhanced the electrochemical response which allowed the development of a rapid (25 min) and sensitive thin layer-based amperometric (TLA) enzyme immunoassay. The method was successfully compared to commercially-available immunofluorescent and real-time PCR assays for RSV testing in respiratory secretion clinical samples. This suggests that owing to its rapidity, convenience, low-cost, portability and ability to provide quantified results, the reported concept could be a promising point-of-care diagnostic tool to screen patients with suspected respiratory infection or other types of infectious diseases.
authors.affiliation: Université de Bourgogne, UMR 1347 Agroécologie, 7 Boulevard Jeanne d'Arc, 21000 Dijon, France. Murielle.Dequaire@u-bourgogne.fr
authors.forename: Murielle
authors.initials: M
authors.lastname: Rochelet
authors.forename: Sébastien
authors.initials: S
authors.lastname: Solanas
authors.forename: Céline
authors.initials: C
authors.lastname: Grossiord
authors.forename: Patricia
authors.initials: P
authors.lastname: Maréchal
authors.forename: Cécile
authors.initials: C
authors.lastname: Résa
authors.forename: Fabienne
authors.initials: F
authors.lastname: Vienney
authors.forename: Come
authors.initials: C
authors.lastname: Barranger
authors.forename: Martine
authors.initials: M
authors.lastname: Joannes
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D015374:Biosensing Techniques / Q000379:methods*; D003124:Colorimetry; D004563:Electrochemistry; D006735:Horseradish Peroxidase / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis*; D012136:Respiratory Syncytial Viruses / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D006735:Horseradish Peroxidase