Evaluation of Elispot assays: Influence of method and operator on variability of results Journal Article


Authors: Janetzki, S.; Schaed, S.; Blachere, N. E.; Ben-Porat, L.; Houghton, A. N.; Panageas, K. S.
Article Title: Evaluation of Elispot assays: Influence of method and operator on variability of results
Abstract: In this study, a comprehensive comparative analysis of different evaluation methods of Elispot plates was performed. Three investigators using three different evaluation approaches read 50 randomly selected wells at three different time points. The methods were: (1) manual evaluation using a stereomicroscope, (2) automated evaluation using an image analysis reader system with reading parameters established by each investigator, and (3) automated evaluation using a reader system with preset reading parameters using assay-specific controls. We demonstrate that manual evaluation had the highest variability both within the same method and when comparing all methods, followed by automated evaluation with investigator-dependent parameters. The variability was low only when all investigators used the same parameters established using assay-specific controls. This variability was independent of operator or spot number per well. Based on this study, recommendations for standardization and validation procedures of Elispot assay performance and evaluation procedures are presented. © 2004 Elsevier B.V. All rights reserved.
Keywords: controlled study; human cell; validation process; reproducibility; reproducibility of results; image analysis; imaging system; standardization; immunoenzyme techniques; evaluation; intermethod comparison; randomization; enzyme linked immunospot assay; stereomicroscopy; microscope; interferon type ii; elispot; laboratory automation; humans; human; priority journal; article; laboratory personnel; elispot plate evaluation
Journal Title: Journal of Immunological Methods
Volume: 291
Issue: 1-2
ISSN: 0022-1759
Publisher: Elsevier Science, Inc.  
Date Published: 2004-08-01
Start Page: 175
End Page: 183
Language: English
DOI: 10.1016/j.jim.2004.06.008
PROVIDER: scopus
PUBMED: 15345315
DOI/URL:
Notes: J. Immunol. Methods -- Cited By (since 1996):26 -- Export Date: 16 June 2014 -- CODEN: JIMMB -- Source: Scopus
Altmetric Score
MSK Authors
  1. Susanne G Schaed
    5 Schaed
  2. Katherine S Panageas
    331 Panageas
  3. Alan N Houghton
    272 Houghton