Quick staining technique for myeloperoxidase using potassium iodide and oxidized pyronine B

Abstract

Myeloperoxidase (MPO) staining has been important for the cytomorphological diagnosis and classification of leukemia. A novel staining method for MPO and its clinical application are presented in the report. Pyronine B (PyB), serving as a chromogenic reagent, was pre-oxidized to obtain stable oxidized Pyronine B solution. The MPO working solution for oxidized pyronine B method consisted of phosphate buffer solution, potassium iodide (KI) solution, and oxidized Pyronine B solution. The positive products of the oxidized Pyronine B method of MPO staining were vibrant red particles located in cytoplasm and the nucleus was stained bluish green. Bone marrow smears from 229 patients with acute leukemia or with grossly normal bone marrow were stained by both oxidized Pyronine B method and the conventional Washburn benzidine staining and a comparison revealed no significant difference in the positive detection rate between the two techniques. The new method eliminates the influence of the varying amount of H2O2 on MPO staining. With this method, the reagents were more stable and the staining procedure was simple and time-saving. This MPO staining technique is a better alternative than the conventional benzidine-based methods.