Optimising automation of a manual enzyme-linked immunosorbent assay
Date
2012-10
Journal Title
Journal ISSN
Volume Title
Publisher
AOSIS Publishing
Abstract
Objective: 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.
Description
CITATION: 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.
The original publication is available at http://www.ajlmonline.org
The original publication is available at http://www.ajlmonline.org
Keywords
Enzyme-linked immunosorbent assay -- Automation, Immunoglobulins, Enzyme-linked immunosorbent assay -- Validity, Infection -- Immunological aspects -- Testing, Vaccination -- Immunological aspects -- Testing
Citation
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