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An international comparability study to determine the sources of uncertainty associated with a fluorescent ELISA

Noble, J E; Wang, L*; Cerasoli, E; Knight, A E; Porter, R A*; Gray, E*; Howe, C*; Hannes, E*; Corbisier, P*; Wang, J*; Wu, L*; Altieri, I*; Patriarca, M*; Hoffman, A*; Resch-Genger, U*; Ebert, B*; Voigt, J*; Shigeri, Y*; Vonsky, M S*; Konopelko, L A*; Gaigalas, A K*; Bailey, M J A (2008) An international comparability study to determine the sources of uncertainty associated with a fluorescent ELISA. Clin. Chem. Lab. Med., 46 (7). pp. 1033-1045.

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Abstract

Background: Immunoassays allow the specific detection of biological molecules in complex samples at physiologically relevant concentrations, however there are concerns over the comparability of such techniques when the same test is performed by different operator, or laboratory. An international intercomparison study was performed to assess the uncertainty involved in the estimation of a protein cytokine concentration using a fluorescent ELISA.
Methods: The laboratories chosen for the study represented national measurement institutes, or nominated expert laboratories. The method was based on a non-competitive sandwich immunoassay with an enhancement step to generate a fluorescent readout. The intercomparison was performed in 2 phases, where the uncertainty of the instrument was determined separately from that of the assay.
Results: Participants were asked to determine an undisclosed concentration of interferon using the supplied standard. The mean participant estimate and SE was 3.54 ± 0.22 mg/ml, with the spread of data ranging ± 35 % of the mean. The quantitation range of the ELISA and of participant's instruments displayed a large variation that contributed to the overall uncertainty.
Conclusions: Sources of uncertainty within the ELISA methodology included; pipetting, model selection and instrument/plate variation.

Item Type: Article
Keywords: ELISA, fluorescence, interferon, uncertainty
Subjects: Biotechnology
Biotechnology > Bio-Diagnostics
Last Modified: 02 Feb 2018 13:15
URI: http://eprintspublications.npl.co.uk/id/eprint/4220

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