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RESEARCH PRODUCT

A thin layer-based amperometric enzyme immunoassay for the rapid and sensitive diagnosis of respiratory syncytial virus infections

Murielle RocheletCécile ResaP. MarechalFabienne VienneyCéline GrossiordSébastien SolanasMartine JoannesC. Barranger

subject

Time Factorsrespiratory syncytial virus[SDV]Life Sciences [q-bio]Biosensing TechniquesRespiratory Syncytial Virus Infectionsscreen-printed sensorSensitivity and SpecificityHorseradish peroxidaseVirusAnalytical ChemistryImmunoenzyme TechniquesElectrochemistrymedicineHumans[SDV.BV]Life Sciences [q-bio]/Vegetal BiologyimmunoassayAntigens ViralHorseradish PeroxidaseChromatographybiologymedicine.diagnostic_testChemistryAntibodies MonoclonalRespiratory infectionSubstrate (chemistry)Molecular biologyAmperometryRespiratory Syncytial VirusesHRP labelImmunoassay[SDE]Environmental Sciencesbiology.proteinColorimetrythin layer amperometric detectionAntibodyConjugate

description

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.

10.1016/j.talanta.2012.07.088https://hal.inrae.fr/hal-02646305