Antigen-induced cytokine and chemokine release test for tuberculosis infection using adsorption of stimulated whole blood on filter paper and multiplex analysis.
Author(s): Skogstrand K, Thysen AH, Jørgensen CS, Rasmussen EM, Andersen AB, Lillebaek T, Hougaard DM, Houen G
Publication: Scand J Clin Lab Invest, 2012, Vol. 72, Page 204-11
PubMed ID: 22283828 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine the effects of conditions of LPS stimulation of blood and preservation of LPS-stimulated blood as DBS or frozen plasma on the levels of 21 inflammatory markers.
Summary of Findings:
Inflammatory markers increased when blood was incubated with LPS, and the rate of increase was larger at 37 degrees C than at room temperature; however, after 24 h a greater change in IFN-gamma was observed at room temperature than when incubated at 37 degrees C. Increases in inflammatory marker levels were generally not found when blood was incubated with saline at room temperature, but small increases were seen after incubation with saline at 37 degrees C. While increases in inflammatory marker levels were noted when specimens were stored, after LPS stimulation, as frozen plasma or as DBS, concentrations of IL-6, 8 and 10 were higher in frozen plasma, and IFN-gamma was higher in DBS. Levels of IL-4, IL-8, IL-18, RANTES, MMP-9, IFN-gamma, IL-1 beta, TNF-beta, MCP-1, and TGF-beta were generally lower, while levels of GM-CSF, IL-2, sIL-6Ra, MIP-1 alpha, IL-17, and TNF-alpha were generally higher in frozen plasma than in DBS from unstimulated blood.
Biospecimens
Preservative Types
- Frozen
- Other Preservative
Diagnoses:
- Not specified
- Normal
- Other diagnoses
Platform:
Analyte Technology Platform Protein ELISA Cell count/volume Lymphocyte proliferation assay Cell count/volume Light microscopy DNA PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Air-dried
Frozen
Preaquisition Diagnosis/ patient condition Tuberculosis
Analyte Extraction and Purification Incubation duration/condition Room temperature
37 degrees C
5 min
2 h
4 h
6 h
24 h
With saline
With LPS
ELISA Specific Targeted peptide/protein IFN gamma
IL-4
IL-6
IL-8
IL-10
IL-18
RANTES
MMP-9
IL-1 beta
TNF-beta
MCP-1
TGF-beta
GM-CSF
IL-2
SIL6ra
MIP-1 alpha
IL-17
TNF-alpha