Utility of a Single-Tube, Six-Color Flow Cytometry Panel for the Diagnosis of Myelodysplastic Syndrome: Experience of a Tertiary Care Centre in India

  • R. Gupta Department of Hematology, Sanjay Gandhi Post Graduate Institute of Medical Sciences Lucknow, India
  • K. Rahman Department of Hematology, Sanjay Gandhi Post Graduate Institute of Medical Sciences Lucknow, India
  • M. K. Singh Department of Hematology, Sanjay Gandhi Post Graduate Institute of Medical Sciences Lucknow, India
  • S. Kumari Department of Hematology, Sanjay Gandhi Post Graduate Institute of Medical Sciences Lucknow, India
  • G. Yadav Department of Hematology, Sanjay Gandhi Post Graduate Institute of Medical Sciences Lucknow, India
  • S. Nityanand Department of Hematology, Sanjay Gandhi Post Graduate Institute of Medical Sciences Lucknow, India
Keywords: MDS, Flow cytometry, FCM score, India

Abstract

Background:  Diagnosis of myelodysplastic syndromes (MDS) is challenging in the presence of morphological mimickers. Flow cytometric immunophenotyping (FCI) has been added to the diagnostic armamentarium, but its use in clinical practice is variable.Materials and Methods: Bone marrow aspirate samples from 54 patients with a clinical and/or morphological suspicion of MDS were subjected to FCI using a single-tube, 6-colour panel comprising of monoclonal antibodies against CD13, CD11b, CD16, CD34, CD45 and CD56. Analysis was centered on the abnormal maturation pattern of granulocytes, blast percentage (≥3%) and ratio of side scatter peak channel value (SSC-PCV) of granulocytes and lymphocytes. Each of these parameters was assigned a score of 1. Overall sensitivity and specificity of this panel was ascertained to differentiate cytopenia/s of MDS from non-MDS cases.Results: Forty MDS and 14 non-MDS cases were diagnosed based on morphology and cytogenetic results. Twenty control samples were also processed simultaneously for FCI to assign the cutoff for various flow cytometric parameters. A score ≥2 was defined as positive for MDS. Hypogranularity was present in 62.5% cases of MDS. The median SSC-PCV of granulocytes and lymphocytes was 6.16 in the MDS group, 7.9 in the non-MDS group and 8.90 in the control group (p <0.05). This cut-off value of 6.16 had a specificity of 92.5% based on the ROC curve analysis. Abnormal granulocyte maturation patterns for CD13/16, CD13/11b and CD11b/16 dot plots were observed in 95.3, 69.8 and 74.4% cases, respectively. The overall sensitivity and specificity of the panel was found to be 87.5% and 64.2%, respectively.Conclusion:  FCI is now an important tool for diagnostic workup in patients presenting with persistent cytopenia with or without morphological evidence of dyspoiesis. Inclusion of objective parameters like SSC-PCV would also reduce inter-lab variability in MDS diagnosis. 

References

Swerdlow S, Harris N, et al. WHO classification of tumors of hematopoietic and lymphoid tissues. Lyon 2008.

Hasse D. Cytogenetic features in myelodysplastic syndromes. Ann Hematol. 2008; 87(7): 515–526.

Solé F, Luño E, Sanzo C et al. Identification Of Novel Cytogenetic Markers With Prognostic Significance In A Series Of 968 Patients With Primary Myelodysplastic Syndromes.Haematologica. 2005;90(9):1168-78.

Stetler-Stevenson M, Arthur DC, Jabbour N, et al. Diagnostic utility of flow cytometric immunophenotyping in myelodysplastic syndrome. Blood 2001;98(4):979-87.

van de Loosdrecht AA, Alhan C, Béné MC, et al. Theresia M. Westers. Standardization of flow cytometry in myelodysplastic syndromes: report from the first European LeukemiaNet working conference on flow cytometry inmyelodysplastic syndromes. Haematologica 2009;94(8):1124-34.

SA W. Diagnosis of myelodysplastic syndromes in cytopenic patients. Hematol Oncol Clin North Am 2011;25(5):1085-110.

Porwit A , Bettelheim P, Brodersen LE, et al. Revisiting guidelines for integration of flow cytometry results in the WHO classification of myelodysplastic syndromes-proposal from the International/European LeukemiaNet Working Group for Flow Cytometry in MDS. Leukemia. 2014;28(9):1793-8.

Van Dongen JJ, Lhermitte L, Böttcher S, et al. EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukemia.2012;26(9):1908-75.

Westers TM IR, Kern W, Alhan C, et al. Standardization of flow cytometry in myelodysplastic syndromes: a report from an international consortium and the European LeukemiaNet Working Group. Leukemia. 2012;26(7):1730-41.

Ogata K. Diagnostic flow cytometry for low-grade myelodysplastic syndromes. Hematol Oncol. 2008;26(4):193-8.

Chopra A, Pati H, Mahapatra M, et al. Flow cytometry in myelodysplastic syndrome: analysis of diagnostic utility using maturation pattern-based and quantitative approaches. Ann Hematol. 2012;91(9):1351-62.

Kern W, Bacher U, Haferlach C, et al. Multiparameter flow cytometry provides independent prognostic information in patients with suspected myelodysplastic syndromes: A study on 804 patients. Cytometry B Clin Cytom. 2015;88(3):154-64.

Della Porta MG LF, Del Vecchio L. Flow cytometry immunophenotyping for the evaluation of bone marrow dysplasia. Cytometry B Clin Cytom. 2011;80(4):201-11

Della Porta MG, Picone C, Pascutto C et al. Multicenter validation of a reproducible flow cytometric score for the diagnosis of low-grade myelodysplastic syndromes: results of a European LeukemiaNET study. Haematologica. 2012;97(8):1209-17.

Chung JW, Park CJ, Cha CH, et al. A combination of CD15/CD10, CD64/CD33, CD16/CD13 or CD11b flow cytometric granulocyte panels is sensitive and specific for diagnosis of myelodysplastic syndrome. Ann Clin Lab Sci. 2012;42(3):271-80.

Published
2018-01-05
How to Cite
1.
Gupta R, Rahman K, Singh M, Kumari S, Yadav G, Nityanand S. Utility of a Single-Tube, Six-Color Flow Cytometry Panel for the Diagnosis of Myelodysplastic Syndrome: Experience of a Tertiary Care Centre in India. ijhoscr. 12(1):29-4.
Section
Original Article(s)