title: A single tube, four-color flow cytometry assay for evaluation of ZAP-70 and CD38 expression in chronic lymphocytic leukemia.
abstract: We describe a simple and robust flow cytometry assay for ZAP-70 and CD38 expression. The steps required to validate this assay in a clinical flow cytometry laboratory are described. Two criteria were used to characterize ZAP-70 expression into positive, negative, and indeterminate categories and applied to 111 cases of chronic lymphocytic leukemia (CLL) resulting in 29.7% positive, 56.8% negative, and 13.5% indeterminate cases. A sensitivity-specificity crossover plot between ZAP-70 and CD38 suggested a cutoff of 12.5% for defining CD38 positivity. ZAP-70+ cases were significantly more likely to be at a higher clinical stage and, together with CD38+ cases, were more likely to have unmutated IgV(H). However, for individual patients, the concordance between these markers was not perfect. It may be necessary to evaluate several prognostic markers simultaneously in CLL, and availability of convenient assays for ZAP-70 and CD38 is desirable for optimal clinical decision making.
authors.affiliation: Department of Pathology, Box 3712, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Nagwa M
authors.initials: NM
authors.lastname: Hassanein
authors.forename: Kathryn R
authors.initials: KR
authors.lastname: Perkinson
authors.forename: Felisa
authors.initials: F
authors.lastname: Alcancia
authors.forename: Barbara K
authors.initials: BK
authors.lastname: Goodman
authors.forename: J Brice
authors.initials: JB
authors.lastname: Weinberg
authors.forename: Anand S
authors.initials: AS
authors.lastname: Lagoo
mesh_terms: D051997:ADP-ribosyl Cyclase 1 / Q000097:blood* / Q000235:genetics; D014408:Biomarkers, Tumor / Q000097:blood; D002869:Chromosome Aberrations; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000032:analysis; D005434:Flow Cytometry / Q000379:methods*; D005786:Gene Expression Regulation; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000097:blood / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000097:blood / Q000235:genetics / Q000473:pathology*; D008562:Membrane Glycoproteins / Q000097:blood* / Q000235:genetics; D009367:Neoplasm Staging; D011237:Predictive Value of Tests; D011379:Prognosis; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000097:blood* / Q000235:genetics
chemical_list: D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D008562:Membrane Glycoproteins; D051746:ZAP-70 Protein-Tyrosine Kinase; C497261:ZAP70 protein, human; C498154:CD38 protein, human; D051997:ADP-ribosyl Cyclase 1