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This Article Full Text Full Text (PDF) Data Supplements All Versions of this Article: clinchem.2005.052639v1 51/12/2341    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (21) Citing Articles via Google Scholar Google Scholar Articles by Döcke, W.-D. Articles by Kunz, D. Search for Related Content PubMed PubMed Citation Articles by Döcke, W.-D. Articles by Kunz, D. Related Collections Clinical Immunology

Monitoring Temporary Immunodepression by Flow Cytometric Measurement of Monocytic HLA-DR Expression: A Multicenter Standardized Study

¹ Institute of Medical Immunology, Charité-Campus Mitte, Humboldt University of Berlin, and University Hospital, Charité Berlin, Berlin, Germany.
² Becton Dickinson, San Diego, CA.
³ Institute of Clinical Chemistry and Pathobiochemistry, University Hospital, Aachen, Germany.
⁴ Department of Aneasthesiology and Intensive Care Medicine, University Hospital, Bonn, Germany.
⁵ Anaesthesia Clinic, University Hospital, Erlangen, Germany.
⁶ Institute of Immunology, University Hospital, Essen, Germany.
⁷ Institute of Clinical Chemistry, University Hospital, Hamburg-Eppendorf, Germany.
⁸ Laboratory of Flowcytometry, University Hospital, Heidelberg, Germany.
⁹ BD Biosciences, Heidelberg, Germany.
¹⁰ Immunology Laboratory, Hopital Neurologique, Lyon South Hospital, Lyon, France.
¹¹ Surgical Research Laboratories, University of Vienna, Vienna, Austria.
¹² Department of Nephrology and Internal Intensive Care, Clinic of Internal Medicine, Charité-Campus Virchow, Humboldt University of Berlin, Berlin, Germany.
¹³ Deutsches Rotes Kreuz Hospital, Neuwied, Germany.

^aAddress correspondence to this author at: Institute of Medical Immunology Charité Berlin, Schumannstrassse 20/21, 10098 Berlin, Germany. Fax 49-30-450-524932; e-mail conny.hoeflich{at}charite.de.

Background: Single-center trials have shown that monocytic HLA-DR is a good marker for monitoring the severity of temporary immunodepression after trauma, major surgery, or sepsis. A new test for measuring monocytic HLA-DR is now available.

Methods: We evaluated a new test reagent set for monocytic HLA-DR expression (BD Quantibrite^TM HLA-DR/Monocyte reagent; Becton Dickinson) in single-laboratory and interlaboratory experiments, assessing preanalytical handling, lyse-no-wash (LNW) vs lyse-wash (LW) values, reference values, and the effect of use of different flow cytometers and different instrument settings on test variance.

Results: For preanalytical handling, EDTA anticoagulation, storage on ice as soon as possible, and staining within 4 h after blood collection gave results comparable to values obtained for samples analyzed immediately after collection (mean increase of 4% in monocytic HLA-DR). Comparison of LNW and LW revealed slightly higher results for LNW (18% higher for LNW compared with LW; r = 0.982). Comparison of different flow cytometers and instrument settings gave CVs <4%, demonstrating the independence of the test from these variables and suggesting that this method qualifies as a standardized test. CV values from the interlaboratory comparison ranged from 15% (blood sample unprocessed before transport) to 25% (stained and fixed before transport).

Conclusions: For the BD Quantibrite HLA-DR/Monocyte test, preanalytical handling is standardized. Single-laboratory results demonstrated the independence of this test from flow cytometer and instrument settings. Interlaboratory results showed greater variance than single-laboratory values. This interlaboratory variance was partly attributable to the influence of transport and can be reduced by optimization of transport conditions.

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