Novel 5-color flow cytometric immunophenotyping protocol for leukemia/lymphoma and its clinical application / 中华检验医学杂志

ABSTRACT

Objective To establish a five-color flow cytometric immunophenotyping protocol and valuate its clinical application in leukemia and lymphoma. Methods Samples from 73 cases of acute leukemia and 30 cases of lymphoma were analyzed using a comprehensive antibody panel with five-color combination. The antibody combination CD7/CD33/CD19/CD13/CD45 and MPO/cCD79a/cCD3/CD117/CD45,composed of different lineage distinctive and lineage sensitive markers of B, T and myeloid cells, were applied to determine the lineage originality of leukemia and lymphoma celia. The markers used in the second step analysis were based on the findings of the first step. Results Five-color compensation can be performed automatically and easily with the advanced digital compensation program. The results showed that the expression ratio of CD19 in B-ALL was 100% (27/27), cCD79, pesitivity was 92.6% (25/27) and CD7,cCD3 was expressed in all T-ALL cases (8/8). The B-ALL could be staged depending on the expression level of CD34 ,CD10 ,cμ,sIgM and the T-ALL could be dearly staged dependent on the expression level of CD2,CD1a,,CD4,CD4,CD8. The expression ratio of cMPO, CD117 was 89. 5% (34/38) and 81.6% (31/38) in AML respectively, however CD7 was also expressed in 26.3% (10/38) of AML cases. Combined with morphology, immanophenotypes could be used for diagnosing AML with subtyping. The CD19/SSC gating can be used for immunophenotyping of B cell lymphoma with differential diagnosis. The number of robes required was significantly reduced with our panel from 12 tubes of 3-color to 6 tubes of 5-color. Implementation of the five-color protocol had resulted in 20% reduction in reagent costs. Conclusions The application of fivecolor staining protocol significantly improve the sensitivity and accuracy of measurement of immunophenotypes of acute leukemia and lymphoma. It reduces the cost and can be widely applied in the clinical laboratory diagnosis.