Myeloperoxidase detection by three-color flow cytometry and by enzyme cytochemistry in the classification of acute leukemia

In the classification of acute leukemia, the presence of myeloperoxidase (MPO) within the leukemic blasts indicates myeloid leukemia. Previous studies compared enzyme cytochemistry (EC) or flow cytometry (FC) with immunocytochemistry in detecting MPO. Our study is the first direct comparison of EC with 3-color FC in a large group of acute leukemias. We studied 26 cases of acute myeloid leukemia (AML) and 4 cases of B-precursor acute lymphoblastic leukemia (B-ALL). Classification was according to the French-American-British criteria. The cells were analyzed for MPO expression by 3-color FC after cell permeabilization followed by staining with anti-MPO antibody. For FC, the blasts were defined by a combination of light scatter characteristics and dim CD45 expression. Concordance between EC and FC was seen in 27 of 30 cases (23/26 AML and all B-ALL), including all AML cases of M1, M2, M3, and M4 subtypes. In 1 of 4 AML-M0 and 2 of 5 AML-M5a cases, FC demonstrated the presence of MPO in 8%, 86%, and 94% blasts; EC detected none. Three-color FC may be more sensitive than routine EC in demonstrating the presence of MPO in AML cases and may offer the advantage of multiparametric analysis.