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

  • Ankur Ahuja
  • Seema Tyagi
  • Tulika Seth
  • Hara Prasad Pati
  • GPS Gahlot
  • Preeti Tripathi
  • Venkatesan Somasundaram
  • Renu Saxena
Original Article


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 .


Acute myeloid leukemia (AML) Myeloperoxidase (MPO) Cytochemistry Immunohistochemistry (IHC) Flow cytometry (FMC) Bone marrow aspirate (BMA) Bone marrow biopsy (BMB) CD34 Blasts 



We acknowledge the support of our patients, as with their support the study had been feasible.

Author Contribution

AA and ST were involved in conceptualization, designing, writing and critical review of the manuscript and were responsible for overall supervision. TS, HPT, GPSG, PT and VS were involved in literature search, writing and manuscript editing. ST is the overall guarantor of the article.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

Ethical Approval

The present study is in compliance with Ethical Standards.


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Copyright information

© Indian Society of Haematology & Transfusion Medicine 2017

Authors and Affiliations

  • Ankur Ahuja
    • 1
  • Seema Tyagi
    • 2
  • Tulika Seth
    • 2
  • Hara Prasad Pati
    • 2
  • GPS Gahlot
    • 1
  • Preeti Tripathi
    • 2
  • Venkatesan Somasundaram
    • 3
  • Renu Saxena
    • 2
  1. 1.Department of Lab Sciences and Molecular MedicineArmy Hospital (Research and Referral)New DelhiIndia
  2. 2.Department of HaematologyAll India Institute of Medical SciencesNew DelhiIndia
  3. 3.Department of PathologyArmed Forces Medical CollegePuneIndia

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