A Comparison of Enzyme-Linked Immunosorbent Assay versus Multiplex Methodology Using an in Vitro Model of Pulmonary Hypertension and InflammationReport as inadecuate




A Comparison of Enzyme-Linked Immunosorbent Assay versus Multiplex Methodology Using an in Vitro Model of Pulmonary Hypertension and Inflammation - Download this document for free, or read online. Document in PDF available to download.

Enzyme-linked immunosorbent assay ELISA is the most widely used method for measuring a single cytokine. Recent developments in cytokine quantification such as multiple arrays measure multiple cytokines simultaneously. Although good correlations between ELISA and multiplex methods have been observed, side by side comparisons are limited. In the present study we hypothesized that ELISA and Luminex techniques are comparable in detecting cytokines in culture medium when pulmonary artery smooth muscle cells PASMC are exposed to stress. Primary human PASMC were cultured in modular chambers and exposed to 21% FiO2 and peak inspiratory and positive end expiratory pressure of 24 and 8 cmH2O respectively, and 95% FiO2. At 24 hours, culture medium was collected and assayed for interleukin-6 IL-6 and IL-8 by quantitative ELISA and by Human Cytokine 25-Plex Panel using a Luminex 200 analyzer. A comparative analysis of agreement between our ELISA and Luminex data was detailed for control and stress conditions using the Bland-Altman plot analysis. Each assay resulted in comparable increased p < 0.001 levels of IL-6 and IL-8 as compared to control in response to oxidative and biophysical stress. The Bland-Altman analysis demonstrated that 95% of the differences between ELISA and Luminex values were within ±1.96 SD from the mean difference indicated by the 95% limits of agreement for the measurements of IL-6 and IL-8. There was no systematic bias as a function of inflammation level. We conclude that in this cell culture model, ELISA and Luminex are comparable in detecting the levels of IL-6 and IL-8 in the culture medium. If measurements of multiple cytokines are demanded and the amount of sample is limited, Luminex multi-analyte profiling technology is accurate and sensitive.

KEYWORDS

Enzyme-Linked Immunosorbent Assay ELISA, Luminex, Pulmonary Artery Smooth Muscle Cells PASMC, Inflammation, Bland-Altman Plot Analysis

Cite this paper

Zhu, Y. , Alapati, D. , Costa, J. , Maduskuie, V. , Fawcett, P. and Shaffer, T. 2014 A Comparison of Enzyme-Linked Immunosorbent Assay versus Multiplex Methodology Using an in Vitro Model of Pulmonary Hypertension and Inflammation. Journal of Biomedical Science and Engineering, 7, 419-426. doi: 10.4236-jbise.2014.77044.





Author: Yan Zhu, Deepthi Alapati, Joanna Costa, Victoria L. Maduskuie, Paul T. Fawcett, Thomas H. Shaffer

Source: http://www.scirp.org/



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