Evaluation of a novel stable whole blood quality control material for lymphocyte subset analysis: results from the UK NEQAS immune monitoring scheme.

The UK NEQAS immune Monitoring Scheme (UK NEQAS) evaluates the performance of laboratories routinely performing T-lymphocyte subset analysis on HIV-infected individuals. The scheme originally issued fresh whole blood, but a significant problem was that of analyte stability, especially 36 h postphlebotomy. To circumvent this problem, we have developed a novel stabilisation procedure that ensures retention of leucocyte light scatter and immunological staining characteristics for up to 300 days. In addition, the stabilised whole blood preparation is fully compatible with flow cytometer technology, incorporating either whole blood lysis or "no wash, no lyse" techniques. The ranges of interlaboratory coefficient of variation for the stabilised material are now tighter than those previously obtained with fresh whole blood. Development of this novel material has enabled overseas laboratories to participate in the UK NEQAS immune Monitoring Scheme and could in the future lead to the production of reference and/or calibration reagents for leucocyte immunophenotyping.

[1]  A. Givan,et al.  Flow Cytometry: First Principles , 1992 .

[2]  A. Landay,et al.  Assessment of Interlaboratory Variability of Immunophenotyping , 1993, Annals of the New York Academy of Sciences.

[3]  P. Altavista,et al.  Quality control study of the Italian group of cytometry on flow cytometry cellular DNA content measurements. , 1993, Cytometry.

[4]  D. Barnett,et al.  Laboratory control of immunocytochemistry , 1989, European journal of haematology.

[5]  A Orfao,et al.  Performance evaluation of the FACSCount System: a dedicated system for clinical cellular analysis. , 1996, Cytometry.

[6]  M R Loken,et al.  Establishing optimal lymphocyte gates for immunophenotyping by flow cytometry. , 1990, Cytometry.

[7]  M. Loken,et al.  Flow Cytometry Principles for Clinical Laboratory Practice: Quality Assurance for Quantitative Immunophenotyping , 1994 .

[8]  J. Nicholson,et al.  CD4 T-lymphocyte determinations on whole blood specimens using a single-tube three-color assay. , 1993, Cytometry.

[9]  B S Edwards,et al.  Comprehensive quality assessment approach for flow cytometric immunophenotyping of human lymphocytes. , 1989, Cytometry.

[10]  J. Fahey,et al.  Results of the flow cytometry ACTG quality control program: analysis and findings. , 1989, Clinical immunology and immunopathology.

[11]  G. Mazzini,et al.  Quality control study of the Italian Group of Cytometry on flow cytometry DNA content measurements: II. Factors affecting inter- and intralaboratory variability. , 1997, Cytometry.

[12]  J. McCoy,et al.  Quality control in flow cytometry for diagnostic pathology: II. A conspectus of reference ranges for lymphocyte immunophenotyping. , 1994, Cytometry.

[13]  R. Atkins,et al.  A comparison of fixatives and immunohistochemical technics for use with monoclonal antibodies to cell surface antigens. , 1982, American journal of clinical pathology.

[14]  A Schwartz,et al.  Development of Clinical Standards for Flow Cytometry , 1993, Annals of the New York Academy of Sciences.

[15]  C. Caldwell.  Preservation of B-cell-associated surface antigens by chemical fixation. , 1994, Cytometry.

[16]  M. Wansbrough-Jones,et al.  Effect of type of haematology analyser on CD4 count , 1992, The Lancet.

[17]  I. Penttilä,et al.  Use of quality control standards in clinical flow cytometry. , 1991, Annals of medicine.

[18]  J. Coon,et al.  Interlaboratory variation in DNA flow cytometry. Results of the College of American Pathologists' Survey. , 1994, Archives of pathology & laboratory medicine.

[19]  D. Murray,et al.  Detection of false DNA aneuploidy and false DNA multiploidy in flow cytometric DNA analysis. , 1993, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.

[20]  R. Atkins,et al.  A comparison of fixatives and immunohistochemical technics for use with monoclonal antibodies to cell surface antigens. , 1982, American journal of clinical pathology.

[21]  K. Bauer,et al.  Quality Control Issues in DNA Content Flow Cytometry , 1993, Annals of the New York Academy of Sciences.

[22]  J. Nicholson.  Immunophenotyping specimens from HIV-infected persons: laboratory guidelines from the Centers for Disease Control and Prevention. , 1994, Cytometry.

[23]  C. Spino,et al.  Standardization of absolute CD4+ lymphocyte counts across laboratories: an evaluation of the Ortho CytoronAbsolute flow cytometry system on normal donors. , 1995, Cytometry.

[24]  M. Macey.  Flow cytometry : clinical applications , 1993 .

[25]  K. Crainic,et al.  Interlaboratory quality assessment of lymphocyte phenotyping. Etalonorme 1990–1992 surveys , 1993, Biology of the cell.