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Evaluation of a novel proximity ligation assay for the sensitive and rapid detection of foot-and-mouth disease virus
Authors:Nordengrahn Ann  Gustafsdottir Sigrun M  Ebert Katja  Reid Scott M  King Donald P  Ferris Nigel P  Brocchi Emiliana  Grazioli Santina  Landegren Ulf  Merza Malik
Institution:Svanova Biotech AB, Uppsala Science Park, S-751 83 Uppsala, Sweden. ann.nordengrahn@svanova.com
Abstract:A novel proximity ligation assay (PLA) using a pan-serotype reactive monoclonal antibody was developed and evaluated for the detection of foot-and-mouth disease virus (FMDV) in clinical samples collected from field cases of disease. The FMDV-specific PLA was found to be 100 times more sensitive for virus detection than the commonly used antigen capture-ELISA (AgELISA). As few as five TCID50 were detected in individual assays, which was comparable with the analytical sensitivity of real-time RT-PCR. Although this assay was capable of detecting diverse isolates from all seven FMDV serotypes, the diagnostic sensitivity of the PLA assay was lower than real-time RT-PCR mainly due to a failure to detect some SAT 1, SAT 2 and SAT 3 FMDV strains. In conclusion, this new PLA format has high analytical sensitivity for the detection of FMDV in clinical samples and may prove valuable as a rapid and simple tool for use in FMD diagnosis.
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