Comparison of Immunohistochemistry, Cytochemistry, and Flow Cytometry in AML for Myeloperoxidase Detection

Abstract

Acute Myeloid Leukemia (AML) as per World Health Organization (WHO 2008) classification is on the basis of the antigenic characterization, enzymes restriction in the neoplastic myeloid cells and the specific translocations/mutations. AML can be assessed and differentiated by flowcytometry (FCM)/immunohistochemistry (IHC)/cytochemistry techniques. Myeloperoxidase (MPO) is an unequivocal marker to differentiate AML from the acute lymphoblastic leukemia. Despite FCM popularity, it has its limitations, in form of 'dry-tap', cost, and inability of being performed by retrospective analysis. IHC, though an old technique has overcome these disadvantages of FCM. Cytochemistry, on the other hand has its own advantages in being cost-effective; technically easy to do while its disadvantages are its inability to be carried out in the old samples, 'dry-tap' conditions in aleukemic leukemia. There has been non-uniformity in the literature among these techniques especially concerning their sensitivity for MPO. A prospective study was done at All India Institute of Medical Sciences New Delhi from 01 July 2014 to 30 Nov 2015 to include 120 diagnosed acute myeloid leukemia cases. Myeloperoxidase stain was done by cytochemistry, immunohistochemistry and flow cytometry and results were compared. There were 28 cases which showed discrepancies. Out of these 28 cases immunohistochemistry showed positivity in majority (22 cases) followed by flow cytometry (14 cases). Therefore it is important to employ more than one technique and IHC must be included for detection of MPO in all suspected cases of AML especially when negative with FCM .

A Graphene Oxide-Based Fluorescent Aptasensor for the Turn-on Detection of CCRF-CEM

“…Therefore, the early diagnosis of leukemia for the treatment and the improvement of the quality of life of patients is essential. At present, the commonly used method for detecting leukemia is taking peripheral blood cells and bone marrow, after that many kinds of analysis [ 6 ], including cell morphology, cytochemistry [ 7 – 9 ], immunophenotype [ 10 , 11 ], immunohistochemical [ 12 , 13 ], and aptamer-based flow cytometry [ 14 , 15 ], have been carried out. These methods can detect leukemia cells, but they still have many shortcomings such as high cost, low sensitivity, and being complicated.…”

Standardised immunophenotypic analysis of myeloperoxidase in acute leukaemia

“…different protocols across participating centres) and limited correlations being reported by others. 4,[11][12][13][14] . Instead, two pairs of controls were selected, which were unfortunately not fully MPO negative or MPO positive.…”

“…relatively complex and expensive). [2][3][4] The UK National External Quality Assessment Service (NEQAS) has shown that standardisation of flow cytometric assays (at least in terms of dyes, clones and sample preparation) is crucial for reproducibility. 5 Nevertheless, to our best knowledge, no de facto standard or fully standardised flow cytometric assay exists for the MPO status.…”

“…minimise intra/inter‐expert and intra/inter‐laboratory variability via standardisation and automation) and disadvantages (e.g. relatively complex and expensive) 2–4 …”

Effect of penehyclidine hydrochloride on IL-6, TNF-α, HIF- 1α and oxidative stress in lung tissue of young rats with acute lung injury

“…The accumulation of a large number of neutrophils in alveolar cavity is one of the characteristic changes in ALI. The activity of myeloperoxidase (MPO), a redox marker enzyme in neutrophils, is used as an index of the accumulation of neutrophils in lung tissues [14]. Malondialdehyde (MDA) is a product of lipid peroxidation in cell membranes.…”