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Received on January 27, 2006
Accepted on April 26, 2006
Molecular Diagnostics and Genetics |
1 Clinical Research Unit, and Department of Infectious Diseases, Copenhagen University Hospital Hvidovre, Hvidovre, Denmark
2 Clinical Research Unit, Copenhagen University Hospital Hvidovre, Hvidovre, Denmark
* To whom correspondence should be addressed. E-mail: kristian.kofoed{at}hh.hosp.dk.
Background: Sepsis is a common and often fatal disease. Because sepsis can be caused by many different organisms, biomarkers that can aid in diagnosing sepsis and monitoring treatment efficacy are highly warranted. New sepsis markers may provide additional information to complement the currently used markers.
Methods: We used a combination of in-house and commercially available multiplex immunoassays based on Luminex® xMAP technology to assay biomarkers of potential interest in EDTA-plasma samples.
Results: A 3-plex assay for soluble urokinase plasminogen activator receptor (suPAR), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), and macrophage migration inhibiting factor (MIF) was developed and validated in-house. This 3-plex assay was added to a commercially available interleukin-1
(IL-1), IL-6, IL-8, granulocyte/macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor-
(TNF-) human cytokine panel. No cross-reactivity was observed when the assays were combined. Correlation between values obtained with the 8-plex, the 5-cytokine panel, the 3 in-house 1-plex assays, and a suPAR ELISA ranged from 0.86 to 0.99. Mean within- and between-run CVs were 8.0% and 11%, respectively. Recoveries of suPAR, sTREM-1, and MIF calibrators were 108%, 88%, and 51%, respectively. In plasma collected from 10 patients with bacterial sepsis confirmed by blood culture, the assay detected significantly increased concentrations of all 8 analytes compared with healthy controls.
Conclusions: A commercially available xMAP panel can be expanded with markers of interest. The combined multiplex assay can measure the 8 analytes with high reproducibility. The xMAP technology is an appealing tool for assaying conventional cytokines in combination with new markers.
The following articles in journals at HighWire Press have cited this article:
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  F. Chergui, A.-S. Chretien, S. Bouali, C. Ramacci, M. Rouyer, T. Bastogne, P. Genin, A. Leroux, and J.-L. Merlin Validation of a Phosphoprotein Array Assay for Characterization of Human Tyrosine Kinase Receptor Downstream Signaling in Breast Cancer Clin. Chem., July 1, 2009; 55(7): 1327 - 1336. [Abstract] [Full Text] [PDF]
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 H.-L. Wong, R. M. Pfeiffer, T. R. Fears, R. Vermeulen, S. Ji, and C. S. Rabkin Reproducibility and Correlations of Multiplex Cytokine Levels in Asymptomatic Persons Cancer Epidemiol. Biomarkers Prev., December 1, 2008; 17(12): 3450 - 3456. [Abstract] [Full Text] [PDF]
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 A.-D. Gu, H.-Y. Mo, Y.-B. Xie, R.-J. Peng, J.-X. Bei, J. Peng, M.-Y. Li, L.-Z. Chen, Q.-S. Feng, W.-H. Jia, et al. Evaluation of a Multianalyte Profiling Assay and an Enzyme-Linked Immunosorbent Assay for Serological Examination of Epstein-Barr Virus-Specific Antibody Responses in Diagnosis of Nasopharyngeal Carcinoma Clin. Vaccine Immunol., November 1, 2008; 15(11): 1684 - 1688. [Abstract] [Full Text] [PDF]
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