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Clinical and Diagnostic Laboratory Immunology, May 2000, p. 344-351, Vol. 7, No. 3
1071-412X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Multisite Comparison of CD4 and CD8 T-Lymphocyte Counting by Single- versus Multiple-Platform Methodologies: Evaluation of Beckman Coulter Flow-Count Fluorospheres and the tetraONE System

Keith A. Reimann,1,* Maurice R. G. O'Gorman,2 John Spritzler,3 Cynthia L. Wilkening,3 Daniel E. Sabath,4 Karen Helm,5 Donald E. Campbell,6 and The NIAID DAIDS New Technologies Evaluation Groupdagger

Division of Viral Pathogenesis, Beth Israel Deaconess Medical Center, Harvard Medical School,1 and Harvard School of Public Health,3 Boston, Massachusetts; Children's Memorial Hospital, Northwestern University, Chicago, Illinois2; University of Washington, Seattle, Washington4; University of Colorado Health Sciences Center, Denver, Colorado5; and Children's Hospital of Philadelphia, Philadelphia, Pennsylvania6

Received 22 February 1999/Returned for modification 23 April 1999/Accepted 18 August 1999

New analytic methods that permit absolute CD4 and CD8 T-cell determinations to be performed entirely on the flow cytometer have the potential for improving assay precision and accuracy. In a multisite trial, we compared two different single-platform assay methods with a predicate two-color assay in which the absolute lymphocyte count was derived by conventional hematology. A two-color method employing lymphocyte light scatter gating and Beckman Coulter Flow-Count fluorospheres for absolute counting produced within-laboratory precision equivalent to that of the two-color predicate method, as measured by coefficient of variation of replicate measurements. The fully automated Beckman Coulter tetraONE System four-color assay employing CD45 lymphocyte gating, automated analysis, and absolute counting by fluorospheres resulted in a small but significant improvement in the within-laboratory precision of CD4 and CD8 cell counts and percentages suggesting that the CD45 lymphocyte gating and automated analysis might have contributed to the improved performance. Both the two-color method employing Flow-Count fluorospheres and the four-color tetraONE System provided significant and substantial improvements in between-laboratory precision of absolute counts. In some laboratories, absolute counts obtained by the single-platform methods showed small but consistent differences relative to the predicate method. Comparison of each laboratory's absolute counts with the five-laboratory median value suggested that these differences resulted from a bias in the absolute lymphocyte count obtained from the hematology instrument in some laboratories. These results demonstrate the potential for single-platform assay methods to improve within-laboratory and between-laboratory precision of CD4 and CD8 T-cell determinations compared with conventional assay methods.

* Corresponding author. Mailing address: Beth Israel Deaconess Medical Center, RE-113, P.O. Box 15732, Boston, MA 02215. Phone: (617) 667-4583. Fax: (617) 667-8210. E-mail: kreimann{at}caregroup.harvard.edu.

dagger M. O'Gorman, Chairman, Northwestern University, Chicago, Ill.; E. Bessent, University of California, San Diego; S. Plaeger, University of California, Los Angeles; A. Donnenberg and S. Douglas, Children's Hospital of Philadelphia, Philadelphia, Pa.; F. Mandy, Bureau for HIV/AIDS and STD, Health Canada, Ottawa, Canada; J. Nicholson, Centers for Disease Control and Prevention, Atlanta, Ga.; K. Reimann, Beth Israel Deaconess Medical Center, Boston, Mass.; J. Schmitz, University of North Carolina; Chapel Hill; C. Schnizlein-Bick; Indiana University, Indianapolis; J. Kagan and D. Livnat, DAIDS/NIAID/National Institutes of Health, Bethesda, Md.

Clinical and Diagnostic Laboratory Immunology, May 2000, p. 344-351, Vol. 7, No. 3
1071-412X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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