Important note: Information in this article was accurate in 2000. The state of the art may have changed since the publication date.
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.The NIAID DAIDS New Technologies Evaluation Group [see comments]
Clin Diagn Lab Immunol. 2000 May;7(3):344-51. Unique Identifier : AIDSLINE MED/20261938 Reimann KA; O'Gorman MR; Spritzler J; Wilkening CL; Sabath DE; Helm K; Campbell DE; Division of Viral Pathogenesis, Beth Israel Deaconess Medical; Center, Harvard Medical School, Boston, MA 02215, USA.; kreimann@caregroup.harvard.edu
Abstract:
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.
Keywords: JOURNAL ARTICLE Antigens, CD45/ANALYSIS Comparative Study CD4-CD8 Ratio/*INSTRUMENTATION/*METHODS Flow Cytometry/*INSTRUMENTATION/*METHODS/STANDARDS Human Immunophenotyping/INSTRUMENTATION/METHODS/STANDARDS Laboratories/STANDARDS Reproducibility of Results Specimen Handling Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S.
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Important note: Information in this article was accurate in 2000. The state of the art may have changed since the publication date.
