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Expression of CD10, CD13, CD19, CD20, CD22, CD33, CD34 and CD45 on B lymphoblasts in bone marrow aspirate of adult patients with newly diagnosed acute lymphoblastic leukemia using multicolor flow cytometry
Authors :
Abstract Background: According to the 2008 WHO classification, the diagnosis of ALL relies on morphological demonstration of presence of 25% or more blast cells in bone marrow in ALL, and to consciously decide on the B lymphoid lineage of the blast cell population. Aim of the study: To evaluate the expression of CD10, CD13, CD19, CD20, CD22, CD33, CD34 and CD45 on B lymphoblasts in bone marrow aspirate of adult patients with newly diagnosed acute lymphoblastic leukemia using multicolor flow cytometry. Patients, materials and methods: This is a cohort study which included 14 newly diagnosed adult patients with B ALL from April 2012 to May 2013. Flow cytometry analysis for CD10, CD13, CD19, CD20, CD22, CD33, CD34 and CD45 was carried out in a private laboratory on bone marrow aspirate samples using 2-laser, 4-color PARTEC Cube6 and using De Novo FCS Express version 4 Flow Cytometry software. The sensitivity of fluorescent detectors was monitored using standard beads according to the manufacturer’s recommendations and normal lymphoid cells within specimens served as internal positive and negative controls. Samples with blasts that aberrantly expressed CD13 or CD33 were tested with SBB cytochemical stain. Results: The range and median percentages for B lymphoblasts, cells in the CD45dim versus low SSC gate, CD10, CD19, CD20, CD22, CD34, CD13 and CD33 were 68-99 and 85, 74-99 and 86, 41-88 and 68, 60-99 and 80, 48-65 and 57, 48-99 and 75, 71-98 and 83, 20-51 and 37, and 20-43 and 30% respectively. Conclusions: FC testing should be accompanied by proper morphological evaluation. For diagnosis of B ALL, start with serial gating using FSC versus SSC, then CD45 versus SSC, then CD34 versus SSC, then using CD19, CD10 and CD22 to confirm the lineage of blasts as B lymphoid.

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Iraqi journal of hematology 2014 ;3(1):47-50