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Clinical and Diagnostic Laboratory Immunology logoLink to Clinical and Diagnostic Laboratory Immunology
. 1997 Mar;4(2):173–179. doi: 10.1128/cdli.4.2.173-179.1997

Inter- and intrainstitutional evaluation of automated volumetric capillary cytometry for the quantitation of CD4- and CD8-positive T lymphocytes in the peripheral blood of persons infected with human immunodeficiency virus. Site Investigators and the NIAID New CD4 Technologies Focus Group.

M R O'Gorman 1, R Gelman 1
PMCID: PMC170497  PMID: 9067651

Abstract

Volumetric capillary cytometry (VCC) is a new technology that involves the detection and enumeration of dually fluorochrome-labeled cells in a precise volume. We compared the accuracy and precision of VCC with the accuracy and precision of flow cytometry and hematology (F&H) for the measurement of the absolute numbers of CD4 and CD8 T cells in the whole blood of patients infected with human immunodeficiency virus. Five laboratories, each with a different F&H system and certified by the National institute of Allergy and Infectious Diseases flow cytometry proficiency testing program, were shipped aliquots of the same samples from a central site in addition to procuring samples locally. In general, the VCC technology generated CD4 and CD8 T-cell counts which were lower than those obtained with F&H. Intralaboratory variability of replicate CD4 T-cell determinations was similar for both technologies except in the local samples with CD4 counts less than 200/microliters, where the VCC variability was higher than the F&H variability. Interlaboratory variability on replicate CD4 T-cell counts made by VCC was significantly less than that when counts were made by F&H. The VCC instrument has automated CD4 and CD8 T-cell enumeration in whole blood and has consolidated the process to a single platform. Its performance in this evaluation indicates that it may represent a viable alternative to F&H for obtaining absolute T-cell subset counts.

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Selected References

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  1. Aboulker J. P., Autran B., Beldjord K., Touraine F., Debre P. Consistency of routine measurements of CD4+, CD8+ peripheral blood lymphocytes. J Immunol Methods. 1992 Oct 2;154(2):155–161. doi: 10.1016/0022-1759(92)90187-x. [DOI] [PubMed] [Google Scholar]
  2. Bentley S. A., Johnson A., Bishop C. A. A parallel evaluation of four automated hematology analyzers. Am J Clin Pathol. 1993 Dec;100(6):626–632. doi: 10.1093/ajcp/100.6.626. [DOI] [PubMed] [Google Scholar]
  3. Bland J. M., Altman D. G. Statistical methods for assessing agreement between two methods of clinical measurement. Lancet. 1986 Feb 8;1(8476):307–310. [PubMed] [Google Scholar]
  4. Connelly M. C., Knight M., Giorgi J. V., Kagan J., Landay A. L., Parker J. W., Page E., Spino C., Wilkening C., Mercolino T. J. Standardization of absolute CD4+ lymphocyte counts across laboratories: an evaluation of the Ortho CytoronAbsolute flow cytometry system on normal donors. Cytometry. 1995 Sep 15;22(3):200–210. doi: 10.1002/cyto.990220307. [DOI] [PubMed] [Google Scholar]
  5. Cordiali Fei P., Solmone M., Vanacore P., Giglio A., Bonifati C., Carducci M., Mussi A., Chiarotti F., Ameglio F. CD4 Lymphocyte enumeration: comparison between flow cytometry and enzyme immunoassay. Cytometry. 1995 Mar 15;22(1):70–74. doi: 10.1002/cyto.990220113. [DOI] [PubMed] [Google Scholar]
  6. Denny T. N., Jensen B. D., Gavin E. I., Louzao A. G., Vella F. A., Oleske J. M., Wong W. Determination of CD4 and CD8 lymphocyte subsets by a new alternative fluorescence immunoassay. Clin Diagn Lab Immunol. 1995 May;2(3):330–336. doi: 10.1128/cdli.2.3.330-336.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dietz L. J., Dubrow R. S., Manian B. S., Sizto N. L. Volumetric capillary cytometry: a new method for absolute cell enumeration. Cytometry. 1996 Mar 1;23(3):177–186. doi: 10.1002/(SICI)1097-0320(19960301)23:3<177::AID-CYTO1>3.0.CO;2-G. [DOI] [PubMed] [Google Scholar]
  8. Fahey J. L., Taylor J. M., Detels R., Hofmann B., Melmed R., Nishanian P., Giorgi J. V. The prognostic value of cellular and serologic markers in infection with human immunodeficiency virus type 1. N Engl J Med. 1990 Jan 18;322(3):166–172. doi: 10.1056/NEJM199001183220305. [DOI] [PubMed] [Google Scholar]
  9. Gelman R., Cheng S. C., Kidd P., Waxdal M., Kagan J. Assessment of the effects of instrumentation, monoclonal antibody, and fluorochrome on flow cytometric immunophenotyping: a report based on 2 years of the NIAID DAIDS flow cytometry quality assessment program. Clin Immunol Immunopathol. 1993 Feb;66(2):150–162. doi: 10.1006/clin.1993.1019. [DOI] [PubMed] [Google Scholar]
  10. Giorgi J. V., Cheng H. L., Margolick J. B., Bauer K. D., Ferbas J., Waxdal M., Schmid I., Hultin L. E., Jackson A. L., Park L. Quality control in the flow cytometric measurement of T-lymphocyte subsets: the multicenter AIDS cohort study experience. The Multicenter AIDS Cohort Study Group. Clin Immunol Immunopathol. 1990 May;55(2):173–186. doi: 10.1016/0090-1229(90)90096-9. [DOI] [PubMed] [Google Scholar]
  11. Jewett J. F., Hecht F. M. Preventive health care for adults with HIV infection. JAMA. 1993 Mar 3;269(9):1144–1153. [PubMed] [Google Scholar]
  12. Johnson D., Hirschkorn D., Busch M. P. Evaluation of four alternative methodologies for determination of absolute CD4+ lymphocyte counts. The National Heart, Lung, and Blood Institute Retrovirus Epidemiology Donor Study. J Acquir Immune Defic Syndr Hum Retrovirol. 1995 Dec 15;10(5):522–530. [PubMed] [Google Scholar]
  13. Koepke J. A., Landay A. L. Precision and accuracy of absolute lymphocyte counts. Clin Immunol Immunopathol. 1989 Jul;52(1):19–27. doi: 10.1016/0090-1229(89)90189-x. [DOI] [PubMed] [Google Scholar]
  14. Landay A., Ho J. L., Hom D., Russell T., Zwerner R., Minuty J. G., Kataaha P., Mmiro F., Jackson B. A rapid manual method for CD4+ T-cell quantitation for use in developing countries. AIDS. 1993 Dec;7(12):1565–1568. doi: 10.1097/00002030-199312000-00004. [DOI] [PubMed] [Google Scholar]
  15. Masur H., Ognibene F. P., Yarchoan R., Shelhamer J. H., Baird B. F., Travis W., Suffredini A. F., Deyton L., Kovacs J. A., Falloon J. CD4 counts as predictors of opportunistic pneumonias in human immunodeficiency virus (HIV) infection. Ann Intern Med. 1989 Aug 1;111(3):223–231. doi: 10.7326/0003-4819-111-3-223. [DOI] [PubMed] [Google Scholar]
  16. Nicholson J. K., Velleca W. M., Jubert S., Green T. A., Bryan L. Evaluation of alternative CD4 technologies for the enumeration of CD4 lymphocytes. J Immunol Methods. 1994 Dec 28;177(1-2):43–54. doi: 10.1016/0022-1759(94)90142-2. [DOI] [PubMed] [Google Scholar]
  17. Paxton H., Pins M., Denton G., McGonigle A. D., Meisner P. S., Phair J. P. Comparison of CD4 cell count by a simple enzyme-linked immunosorbent assay using the TRAx CD4 test kit and by flow cytometry and hematology. Clin Diagn Lab Immunol. 1995 Jan;2(1):104–114. doi: 10.1128/cdli.2.1.104-114.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rickman W. J., Monical C., Waxdal M. J. Improved precision in the enumeration of absolute numbers of lymphocyte phenotypes with long-term monthly proficiency testing. Ann N Y Acad Sci. 1993 Mar 20;677:53–58. doi: 10.1111/j.1749-6632.1993.tb38764.x. [DOI] [PubMed] [Google Scholar]
  19. Robinson G., Morgan L., Evans M., McDermott S., Pereira S., Wansbrough-Jones M., Griffin G. Effect of type of haematology analyser on CD4 count. Lancet. 1992 Aug 22;340(8817):485–485. doi: 10.1016/0140-6736(92)91807-k. [DOI] [PubMed] [Google Scholar]
  20. Schenker E. L., Hultin L. E., Bauer K. D., Ferbas J., Margolick J. B., Giorgi J. V. Evaluation of a dual-color flow cytometry immunophenotyping panel in a multicenter quality assurance program. Cytometry. 1993;14(3):307–317. doi: 10.1002/cyto.990140311. [DOI] [PubMed] [Google Scholar]

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