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Optimising automation of a manual enzyme-linked immunosorbent assay

dc.contributor.authorDe Beer, Corenaen_ZA
dc.contributor.authorEsser, Monikaen_ZA
dc.contributor.authorPreiser, Wolfgangen_ZA
dc.date.accessioned2015-03-11T09:29:43Z
dc.date.available2015-03-11T09:29:43Z
dc.date.issued2012-10
dc.identifier.citationDe Beer, C., Esser, M. & Preiser, W. 2012. Optimising automation of a manual enzyme-linked immunosorbent assay. African Journal of Laboratory Medicine, 1(1): 1-3, doi: 10.4102/ajlm.v1i1.15
dc.identifier.issn2225-2010 (online)
dc.identifier.issn2225-2002 (print)
dc.identifier.otherdoi: 10.4102/ajlm.v1i1.15
dc.identifier.urihttp://hdl.handle.net/10019.1/96254
dc.descriptionCITATION: De Beer, C., Esser, M. & Preiser, W. 2012. Optimising automation of a manual enzyme-linked immunosorbent assay. African Journal of Laboratory Medicine, 1(1): 1-3, doi: 10.4102/ajlm.v1i1.15.
dc.descriptionThe original publication is available at http://www.ajlmonline.org
dc.description.abstractObjective: Enzyme-linked immunosorbent assays (ELISAs) are widely used to quantify immunoglobulin levels induced by infection or vaccination. Compared to conventional manual assays, automated ELISA systems offer more accurate and reproducible results, faster turnaround times and cost effectiveness due to the use of multianalyte reagents. Design: The VaccZyme™ Human Anti-Haemophilus influenzae type B (Hib) kit (MK016) from The Binding Site Company was optimised to be used on an automated BioRad PhDTM system in the Immunology Laboratory (National Health Laboratory Service) in Tygerberg, South Africa. Methods: An automated ELISA system that uses individual well incubation was compared to a manual method that uses whole-plate incubation. Results: Results were calculated from calibration curves constructed with each assay. Marked differences in calibration curves were observed for the two methods. The automated method produced lower-than-recommended optical density values and resulted in invalid calibration curves and diagnostic results. A comparison of the individual steps of the two methods showed a difference of 10 minutes per incubation cycle. All incubation steps of the automated method were subsequently increased from 30 minutes to 40 minutes. Several comparative assays were performed according to the amended protocol and all calibration curves obtained were valid. Calibrators and controls were also included as samples in different positions and orders on the plate and all results were valid. Conclusion: Proper validation is vital before converting manual ELISA assays to automated or semi-automated methods.en_ZA
dc.description.urihttp://www.ajlmonline.org/index.php/ajlm/article/view/15
dc.format.extent3 pages
dc.language.isoen_ZAen_ZA
dc.publisherAOSIS Publishing
dc.subjectEnzyme-linked immunosorbent assay -- Automationen_ZA
dc.subjectImmunoglobulinsen_ZA
dc.subjectEnzyme-linked immunosorbent assay -- Validityen_ZA
dc.subjectInfection -- Immunological aspects -- Testingen_ZA
dc.subjectVaccination -- Immunological aspects -- Testingen_ZA
dc.titleOptimising automation of a manual enzyme-linked immunosorbent assayen_ZA
dc.typeArticleen_ZA
dc.description.versionPublisher's version
dc.rights.holderAuthors retain copyright


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