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Published in: Indian Journal of Hematology and Blood Transfusion 2/2018

01-04-2018 | Original Article

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

Authors: Ankur Ahuja, Seema Tyagi, Tulika Seth, Hara Prasad Pati, GPS Gahlot, Preeti Tripathi, Venkatesan Somasundaram, Renu Saxena

Published in: Indian Journal of Hematology and Blood Transfusion | Issue 2/2018

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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 .
Literature
1.
go back to reference Neame PB, Soamboonsrup P, Browman GP et al (1986) Classifying acute leukemia by immunophenotyping: a combined FAB immunologic classification of acute myelogenous leukemia. Blood 68:1355–1362PubMed Neame PB, Soamboonsrup P, Browman GP et al (1986) Classifying acute leukemia by immunophenotyping: a combined FAB immunologic classification of acute myelogenous leukemia. Blood 68:1355–1362PubMed
2.
go back to reference Bene MC, Castoldi G, Knapp W et al (1995) Proposals for the immunological classification of acute leukemia’s. European group for the immunological characterization of leukemia’s (EGIL). Leukemia 9:1783–1786PubMed Bene MC, Castoldi G, Knapp W et al (1995) Proposals for the immunological classification of acute leukemia’s. European group for the immunological characterization of leukemia’s (EGIL). Leukemia 9:1783–1786PubMed
3.
go back to reference Subashchandrabose P, Madanagopaal LR, Rao TMS (2016) Diagnosis and Classification of Acute Leukemia in Bone Marrow Trephine Biopsies, Utility of a Selected Panel of Minimal Immunohistochemical Markers. IJHOSCR 10(3):138–146PubMedPubMedCentral Subashchandrabose P, Madanagopaal LR, Rao TMS (2016) Diagnosis and Classification of Acute Leukemia in Bone Marrow Trephine Biopsies, Utility of a Selected Panel of Minimal Immunohistochemical Markers. IJHOSCR 10(3):138–146PubMedPubMedCentral
4.
go back to reference Peffault de Latour R, Legrand O, Moreau D, Perrot J, Blanc C, Chaoui D, Casadeall N, Marie J (2003) Comparison of flow cytometry and enzyme cytochemistry for the detection of myeloperoxidase in acute myeloid leukaemia: interests of a new positivity threshold. Br J Haematol 122:211–216CrossRefPubMed Peffault de Latour R, Legrand O, Moreau D, Perrot J, Blanc C, Chaoui D, Casadeall N, Marie J (2003) Comparison of flow cytometry and enzyme cytochemistry for the detection of myeloperoxidase in acute myeloid leukaemia: interests of a new positivity threshold. Br J Haematol 122:211–216CrossRefPubMed
5.
go back to reference Kheiri S, MacKerrell T, Bonagura V, Fuchs A, Billett H (1998) Flow cytometry with or without cytochemistry for the diagnosis of acute leukemia’s. Cytometry 34:82–86CrossRefPubMed Kheiri S, MacKerrell T, Bonagura V, Fuchs A, Billett H (1998) Flow cytometry with or without cytochemistry for the diagnosis of acute leukemia’s. Cytometry 34:82–86CrossRefPubMed
6.
go back to reference Swerdlow SH, Campo E, Harris NL et al (2008) WHO classification of tumours of haematopoietic and lymphoid tissues. IARC Press, Lyon Swerdlow SH, Campo E, Harris NL et al (2008) WHO classification of tumours of haematopoietic and lymphoid tissues. IARC Press, Lyon
7.
go back to reference Saravanan L, Juneja S (2010) Immunohistochemistry is a more sensitive marker for the detection of myeloperoxidase in acute myeloid leukemia compared with flow cytometry and cytochemistry. Int J Lab Hematol 32:132–136CrossRef Saravanan L, Juneja S (2010) Immunohistochemistry is a more sensitive marker for the detection of myeloperoxidase in acute myeloid leukemia compared with flow cytometry and cytochemistry. Int J Lab Hematol 32:132–136CrossRef
8.
go back to reference General Hematology Taskforce of BCSH (1994) Immunophenotyping in the diagnosis of acute leukemia’s. J Clin Pathol 47:777–781CrossRef General Hematology Taskforce of BCSH (1994) Immunophenotyping in the diagnosis of acute leukemia’s. J Clin Pathol 47:777–781CrossRef
9.
go back to reference Prabhu M, Vandana P, Venkatesan S, Abhishek P, Rahul KS, Mandeep D, Renu S (2015) Can threshold for MPO by flow cytometry be reduced in classifying acute leukaemia? A comparison of flow cytometric and cytochemical myeloperoxidase using different flow cytometric cut-offs. Hematology 20:455–461CrossRef Prabhu M, Vandana P, Venkatesan S, Abhishek P, Rahul KS, Mandeep D, Renu S (2015) Can threshold for MPO by flow cytometry be reduced in classifying acute leukaemia? A comparison of flow cytometric and cytochemical myeloperoxidase using different flow cytometric cut-offs. Hematology 20:455–461CrossRef
10.
go back to reference Nakase K, Sartor M, Bradstock K (1998) Detection of myeloperoxidase by flow cytometry in acute leukemia. Cytometry 34:198–202CrossRefPubMed Nakase K, Sartor M, Bradstock K (1998) Detection of myeloperoxidase by flow cytometry in acute leukemia. Cytometry 34:198–202CrossRefPubMed
11.
go back to reference Arber A, Snyder D, Fine M, Dagis A, Niland J, Slovak M (2001) Myeloperoxidase immunoreactivity in adult acute lymphoblastic leukemia. Am J Clin Pathol 116:25–33CrossRefPubMed Arber A, Snyder D, Fine M, Dagis A, Niland J, Slovak M (2001) Myeloperoxidase immunoreactivity in adult acute lymphoblastic leukemia. Am J Clin Pathol 116:25–33CrossRefPubMed
12.
go back to reference Nguyen P, Olszak L, Harris N, Preffer F (1998) Myeloperoxidase detection by three-color flow cytometry and by enzyme cytochemistry in the classification of acute leukemia. Am J Clin Pathol 110:163–169CrossRefPubMed Nguyen P, Olszak L, Harris N, Preffer F (1998) Myeloperoxidase detection by three-color flow cytometry and by enzyme cytochemistry in the classification of acute leukemia. Am J Clin Pathol 110:163–169CrossRefPubMed
13.
go back to reference Lepelley P, Preudhomme C, Sartiaux C, Ghevaert C, Lai J, Iaru T, Fenaux P, Cosson A (1993) Immunological detection of myeloperoxidase in poorly differentiated acute leukemia. Eur J Haematol 50:155–159CrossRefPubMed Lepelley P, Preudhomme C, Sartiaux C, Ghevaert C, Lai J, Iaru T, Fenaux P, Cosson A (1993) Immunological detection of myeloperoxidase in poorly differentiated acute leukemia. Eur J Haematol 50:155–159CrossRefPubMed
14.
go back to reference Van der Schoot C, Daams G, Pinkster J, von Vet R, dem Borne A (1990) Monoclonal antibodies against myeloperoxidase are valuable immunological reagents for the diagnosis of acute myeloid leukemia. Br J Haematol 74:173–178CrossRefPubMed Van der Schoot C, Daams G, Pinkster J, von Vet R, dem Borne A (1990) Monoclonal antibodies against myeloperoxidase are valuable immunological reagents for the diagnosis of acute myeloid leukemia. Br J Haematol 74:173–178CrossRefPubMed
15.
go back to reference Bain B, Barnett D, Linch D, Matutes E, Reilly J (2002) Revised guideline on immunophenotyping in acute leukaemias and chronic lymphoproliferative disorders. Clin Lab Hematol 24:1CrossRef Bain B, Barnett D, Linch D, Matutes E, Reilly J (2002) Revised guideline on immunophenotyping in acute leukaemias and chronic lymphoproliferative disorders. Clin Lab Hematol 24:1CrossRef
16.
go back to reference van den Ancker W, Westers TM, de Leeuw DC, van der Veeken YF, Loonen A, van Beckhoven E, Ossenkoppele GJ, van de Loosdrecht AA (2013) A threshold of 10% for myeloperoxidase by flow cytometry is valid to classify acute leukemia of ambiguous and myeloid origin. Cytom B Clin Cytom 84B:114–118CrossRef van den Ancker W, Westers TM, de Leeuw DC, van der Veeken YF, Loonen A, van Beckhoven E, Ossenkoppele GJ, van de Loosdrecht AA (2013) A threshold of 10% for myeloperoxidase by flow cytometry is valid to classify acute leukemia of ambiguous and myeloid origin. Cytom B Clin Cytom 84B:114–118CrossRef
17.
go back to reference Jaffe E, Harris N, Stein H, Vardiman J (eds) (2001) World health organization classification of tumors. Pathology and genetics of tumors of haematopoietic and lymphoid tissues, chapter 4. IARC Press, Lyon Jaffe E, Harris N, Stein H, Vardiman J (eds) (2001) World health organization classification of tumors. Pathology and genetics of tumors of haematopoietic and lymphoid tissues, chapter 4. IARC Press, Lyon
18.
go back to reference Guy J, Antony-Debré I, Benayoun E, Arnoux I, Fossat C, Le Garff-Tavernier M, Raimbault A et al (2013) Flow cytometry thresholds of myeloperoxidase detection to discriminate between acute lymphoblastic or myeloblasticleukaemia. Br J Haematol 161:551–555CrossRefPubMed Guy J, Antony-Debré I, Benayoun E, Arnoux I, Fossat C, Le Garff-Tavernier M, Raimbault A et al (2013) Flow cytometry thresholds of myeloperoxidase detection to discriminate between acute lymphoblastic or myeloblasticleukaemia. Br J Haematol 161:551–555CrossRefPubMed
Metadata
Title
Comparison of Immunohistochemistry, Cytochemistry, and Flow Cytometry in AML for Myeloperoxidase Detection
Authors
Ankur Ahuja
Seema Tyagi
Tulika Seth
Hara Prasad Pati
GPS Gahlot
Preeti Tripathi
Venkatesan Somasundaram
Renu Saxena
Publication date
01-04-2018
Publisher
Springer India
Published in
Indian Journal of Hematology and Blood Transfusion / Issue 2/2018
Print ISSN: 0971-4502
Electronic ISSN: 0974-0449
DOI
https://doi.org/10.1007/s12288-017-0849-1

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