TITLE:
Introducing a Rapid and Safe Method for Myeloperoxidase Staining
AUTHORS:
Fatemeh E. Mahjoub, Fahimeh Firouzjaie Karder, Issa Jahanzad, Saghi Vaziri, Ramezan Ali Sharifian, Zahra Farahani
KEYWORDS:
Myeloperoxidase Staining, DAB 3, 3-Diaminobenzidine, Safe and Rapid Method
JOURNAL NAME:
Open Journal of Pathology,
Vol.5 No.2,
March
31,
2015
ABSTRACT: Background: Myeloperoxidase staining is
used to differentiate leukemias since several decades. Despite implementation
of flow cytometric, cytogenetic and molecular techniques for identification of
leukemic blasts, histochemical stains such as myeloperoxidase stain are
persistently used for better classification of leukemias. The myeloperoxidase
staining is a time consuming and hazardous procedure. The present report
describes a sensitive, rapid and easy method for assessment of peroxidase
activity. Materials and Methods: Bone marrow aspiration slides were stained
with Dako product: Code number: K3467 containing DAB chromogen (3,3-diaminobenzidine
in chromogen solution) and substrate buffer (Imidasole-HCL buffer, PH 7.5
containing hydrogen peroxide and an anti microbial agent) in a rapid procedure
taking only ten minutes time. The staining needs no material preparation steps.
Neutrophils in the slide are taken as positive control or another normal smear
was costained to be used as control. All cases were followed up with flow
cytometry and cytogenetic studies. Result: The reaction product of this stain
is brown and granular. Promyelocytes and myelocytes are the most strongly
staining cells with positive (primary) granules. Lymphoblasts are negative. The
result of classification of leukemias with this technique was in concordance
with flow cytometric immunophenotyping. Discussion: Many practical techniques
have been described using benzidine as an indicator for myeloperoxidase
staining. Benzidine is a carcinogenic material and its usage is severely
restricted in laboratory. Formerly we prepared requisite materials for
myeloperoxidase staining by hazardous ways (boiling), but we decided to apply
ready to use 3,3-diaminobenzidine (DAB), which is used in final step of
immunohistochemistry stains. Conclusion: Use of 3,3-diaminobenzidine (DAB) is
highly recommended for myeloperoxidase staining, while the result is
extraordinary and fully compatible with flow cytometry and the method is safe
and rapid.