R. Stebbings, L. Wang, J. Sutherland, M. Kammel, A.K. Gaigalas, M. John, B. Roemer, Maren Kuhne, Rudolf Schneider, M. Braun, A. Engel, D.K. Dikshit, F. Abbasi, G.E. Marti, M.P. Sassi, L. Revel, S.-K. Kim, M.-O. Baradez, T. Lekishvili, D. Marshall, L. Whitby, W. Jing, V. Ost, M. Vonsky, J. Neukammer
- A surface-labeled lyophilized lymphocyte (sLL) preparation has been developed using human peripheral blood mononuclear cells prelabeled with a fluorescein isothiocyanate conjugated anti-CD4 monoclonal antibody. The sLL preparation is intended to be used as a reference material for CD4+ cell counting including the development of higher order reference measurement procedures and has been evaluated in the pilot study CCQM-P102. This study was conducted across 16 laboratories from eight countries to assess the ability of participants to quantify the CD4+ cell count of this reference material and to document cross-laboratory variability plus associated measurement uncertainties. Twelve different flow cytometer platforms were evaluated using a standard protocol that included calibration beads used to obtain quantitative measurements of CD4+ T cell counts. There was good overall cross-platform and counting method agreement with a grand mean of the laboratory calculated means of (301.7 ± 4.9)A surface-labeled lyophilized lymphocyte (sLL) preparation has been developed using human peripheral blood mononuclear cells prelabeled with a fluorescein isothiocyanate conjugated anti-CD4 monoclonal antibody. The sLL preparation is intended to be used as a reference material for CD4+ cell counting including the development of higher order reference measurement procedures and has been evaluated in the pilot study CCQM-P102. This study was conducted across 16 laboratories from eight countries to assess the ability of participants to quantify the CD4+ cell count of this reference material and to document cross-laboratory variability plus associated measurement uncertainties. Twelve different flow cytometer platforms were evaluated using a standard protocol that included calibration beads used to obtain quantitative measurements of CD4+ T cell counts. There was good overall cross-platform and counting method agreement with a grand mean of the laboratory calculated means of (301.7 ± 4.9) µL-1 CD4+ cells. Excluding outliers, greater than 90% of participant data agreed within ±15%. A major contribution to variation of sLL CD4+ cell counts was tube to tube variation of the calibration beads, amounting to an uncertainty of 3.6%. Variation due to preparative steps equated to an uncertainty of 2.6%. There was no reduction in variability when data files were centrally reanalyzed. Remaining variation was attributed to instrument specific differences. CD4+ cell counts obtained in CCQM-P102 are in excellent agreement and show the robustness of both the measurements and the data analysis and hence the suitability of sLL as a reference material for interlaboratory comparisons and external quality assessment.…
MetadatenAutor*innen: | R. Stebbings, L. Wang, J. Sutherland, M. Kammel, A.K. Gaigalas, M. John, B. Roemer, Maren Kuhne, Rudolf SchneiderORCiD, M. Braun, A. Engel, D.K. Dikshit, F. Abbasi, G.E. Marti, M.P. Sassi, L. Revel, S.-K. Kim, M.-O. Baradez, T. Lekishvili, D. Marshall, L. Whitby, W. Jing, V. Ost, M. Vonsky, J. Neukammer |
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Dokumenttyp: | Zeitschriftenartikel |
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Veröffentlichungsform: | Verlagsliteratur |
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Sprache: | Englisch |
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Titel des übergeordneten Werkes (Englisch): | Cytometry / A |
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Jahr der Erstveröffentlichung: | 2015 |
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Verlag: | Wiley-Liss |
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Verlagsort: | Hoboken, NJ |
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Jahrgang/Band: | 87 |
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Ausgabe/Heft: | 3 |
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Erste Seite: | 244 |
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Letzte Seite: | 253 |
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Freie Schlagwörter: | Acquired immunodeficiency syndrome; CD4+ cell counting; Flow cytometry; Human immunodeficiency virus-1; Lyophilized cells; Measurement of uncertainty; Reference material; Relative concentration measurement; Standard measurement procedure |
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DOI: | 10.1002/cyto.a.22614 |
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ISSN: | 0196-4763 |
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ISSN: | 1552-4922 |
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ISSN: | 1552-4930 |
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Verfügbarkeit des Dokuments: | Datei im Netzwerk der BAM verfügbar ("Closed Access") |
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Datum der Freischaltung: | 20.02.2016 |
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Referierte Publikation: | Nein |
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