Skip to main content
SpringerLink
Log in

A Prospective, Cross Sectional Study of PNH Clone in MDS Patients Using High Sensitivity Flowcytometry: A Single Center Experience

  • Original Article
  • Published:
Indian Journal of Hematology and Blood Transfusion Aims and scope Submit manuscript

Abstract

Subclinical PNH can be present in patients with bone marrow failure like aplastic anemia and myelodysplastic syndrome (MDS). Such clone may have prognostic and therapeutic implications. In literature around 1–10% MDS cases have shown a PNH clone, however, data from India is relatively scarce. A high sensitivity PNH assay was employed using a single tube combination of FLAER, CD157, CD64, CD15 and CD45 antibodies in adult patients of MDS at presentation. A clone size of  > 0.01% was taken as significant. A total of 30 patients were included. PNH clone was present in 30% cases. Correlation done between PNH clone size and LDH values showed moderately positive correlation (r = 0.735, p = 0.001, r2 = 0.541). As per this study a LDH cut off of 247 IU is likely to predict a PNH clone (> 1%) with moderate sensitivity and specificity. High sensitivity PNH assay is able to detect small PNH clone. Calculating the cut-off of LDH to predict PNH positivity can help us judiciously prescribe this test in MDS patients in resource constrained settings.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Bessler M, Mason PJ, Hillmen P, Miyata T, Yamada N, Takeda N, Luzzatto L, Kinoshita T (1994) Paroxysmal nocturnal hemoglobinuria (PNH) is caused by somatic mutations in the PIG-A gene. EMBO J 13:110–117

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Borowitz MJ, Craig FE, Digiuseppe JA, Illingworth AJ, Rosse W, Sutherland DR, Wittwer CT, Richards SJ (2010) Guidelines for the diagnosis and monitoring of paroxysmal nocturnal hemoglobinuria and related disorders by flow cytometry. Cytom B Clin Cytom 78:211–230

    Google Scholar 

  3. Wang SA, Pozdnyakova O, Jorgensen JL, Medeiros LJ, Stachurski D, Anderson M et al (2009) Detection of paroxysmal nocturnal hemoglobinuria clones in patients with myelodysplastic syndromes and related bone marrow diseases, with emphasis on diagnostic pitfalls and caveats. Haematologica 94:29–36

    Article  PubMed  Google Scholar 

  4. Scheinberg P, Marte M, Nunez O, Young NS (2010) Paroxysmal nocturnal hemoglobinuria clones in severe aplastic anemia patients treated with horse anti-thymocyte globulin plus cyclosporine. Haematologica 95:1075–1080

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Young NS (2009) Paroxysmal nocturnal hemoglobinuria and myelodysplastic sydromes: clonal expansion of PIG-A-mutant hematopoietic cells in bone marrow failure. Haematologica 94(1):3–7

    Article  PubMed  PubMed Central  Google Scholar 

  6. Brodsky RA (2017) Paroxysmal nocturnal hemoglobinuria. In: Hoffman R, Benz EJ, Silberstein LE, Heslop HE, Anastasi J, Abutalib SA et al (eds) Hematology: basic principles and practice, 7th edn. Elsevier, Philadelphia, p 417

    Google Scholar 

  7. Chopra A, Pati H, Mahapatra M, Mishra P, Seth T, Kumar S et al (2012) Flow cytometry in myelodysplastic syndrome: analysis of diagnostic utility using maturation pattern-based and quantitative approaches. Ann Hematol 91(9):1351–1362

    Article  PubMed  Google Scholar 

  8. Shah NM, Prajapati SG, Adesara RP, Patel AP (2009) An analysis of 30 cases of myelodysplastic syndrome. Indian J Pathol Microbiol 52(2):206–209

    Article  PubMed  Google Scholar 

  9. Vundinti BR, Kerketta L, Jijina F, Ghosh K (2009) Cytogenetic study of myelodysplastic syndrome from India. Indian J Med Res 130(2):155–159

    PubMed  Google Scholar 

  10. Aul C, Gattermann N, Schneider W (1992) Age-related incidence and other epidemiological aspects of myelodysplastic syndromes. Br J Haematol 82(2):358–367

    Article  CAS  PubMed  Google Scholar 

  11. Chatterjee T, Dixit A, Mohapatra M, Tyagi S, Gupta PK, Mishra P et al (2004) Clinical, haematological and histomorphological profile of adult myelodysplastic syndrome. Study of 96 cases in a single institute. Eur J Haematol 73(2):93–97

    Article  CAS  PubMed  Google Scholar 

  12. Raza A, Ravandi F, Rastogi A, Bubis J, Lim SH, Weitz I, Castro-Malaspina H, Galili N, Jawde RA, Illingworth A (2014) A prospective multicenter study of paroxysmal nocturnal hemoglobinuria cells in patients with bone marrow failure. Cytom B Clin Cytom 86:175–182

    Article  Google Scholar 

  13. Agarwal R, Chapple P, Brown M, Szer J, Juneja S (2015) Analysis of abnormal clones by the fluorescent aerolysin method in paroxysmal nocturnal haemoglobinuria and other marrow disorders. Int J Lab Hematol 37:14–21

    Article  CAS  PubMed  Google Scholar 

  14. Wang H, Chuhjo T, Yasue S, Omine M, Nakao S (2002) Clinical significance of a minor population of paroxysmal nocturnal hemoglobinuria-type cells in bone marrow failure syndrome. Blood 100:3897–3902

    Article  CAS  PubMed  Google Scholar 

  15. Vundinti BR, Kerketta L, Jijina F, Ghosh K (2009) Cytogenetic study of myelodysplastic syndrome from India. Indian J Med Res 130(2):155–159

    PubMed  Google Scholar 

  16. Chaubey R, Sazawal S, Dada R, Mahapatra M, Saxena R (2011) Cytogenetic profile of Indian patients with de novo myelodysplastic syndromes. Indian J Med Res 134:452–457

    PubMed  PubMed Central  Google Scholar 

  17. Ogata K (2008) Diagnostic flow cytometry for low-grade myelodysplastic syndromes. Hematol Oncol 26(4):193–198

    Article  PubMed  Google Scholar 

  18. Pozdnyakova O, Miron PM, Tang G, Walter O, Raza A, Woda B et al (2008) Cytogenetic abnormalities in a series of 1029 patients with primary myelodysplastic syndromes: a report from the US with a focus on some undefined single chromosomal abnormalities. Cancer 113(12):3331–3340

    Article  PubMed  Google Scholar 

  19. Haase D, Germing U, Schanz J, Pfeilstocker M, Nosslinger T, Hildebrandt B et al (2007) New insights into the prognostic impact of the karyotype in MDS and correlation with subtypes: evidence from a core dataset of 2124 patients. Blood 110(13):4385–4395

    Article  CAS  PubMed  Google Scholar 

  20. Shih A, Chin-Yee IH, Hedley B, Keeney M, Wells RA, Sutherland DR, Hsia CC (2011) Screening patients with myelodysplastic syndrome and aplastic anemia for paroxysmal nocturnal hemoglobinuria clones: a retrospective study. Blood 118(21):3426

    Article  Google Scholar 

  21. Illingworth A, Marinov I, Sutherland DR, Wagner-Ballon O, DelVecchio L (2018) ICCS/ESCCA consensus guidelines to detect GPI-deficient cells in paroxysmal nocturnal hemoglobinuria (PNH) and related disorders part 3—data analysis, reporting and case studies. Cytom Part B 94B:49–66

    Article  Google Scholar 

  22. Sutherland DR, Keeney M, Illingworth A (2012) Practical guidelines for the high sensitivity detection and monitoring of paroxysmal nocturnal hemoglobinuria clones by flow cytometry. Cytom B Clin Cytom 82(4):195–208

    Article  Google Scholar 

  23. Dunn DE, Tanawattanacharoen P, Boccuni P et al (1999) Paroxysmal nocturnal hemoglobinuria cells in patients with bone marrow failure syndromes. Ann Intern Med 131:401–408

    Article  CAS  PubMed  Google Scholar 

  24. Rocha JMC, Silva ML, Souza MEL, Murao M, Araújo SSDS, Santos SME (2014) Detection of PNH cells by flow cytometry, using multiparameter analysis. J Bras Pathol Med Lab 50:105–114

    CAS  Google Scholar 

  25. Rocha JMC, Silva ML, Souza MEL, Murao M, Araújo SSDS, Santos SME (2014) Detection of PNH cells by flow cytometry, using multiparameter analysis. J Bras Patol Med Lab 50:105–114

    Article  CAS  Google Scholar 

  26. Gupta SK, Pati HP, Tejomurtula AP, Seth T (2010) PNH clone assessment by flow cytometry and its clinical correlation in PNH and aplastic anemia. J Hematopathol 3(4):137–143

    Article  Google Scholar 

  27. Rahman K, Gupta R, Yadav G, Husein N, Singh MK, Nityanand S (2017) Fluorescent aerolysin (FLAER)-based paroxysmal nocturnal hemoglobinuria (PNH) screening: a single center experience from India. Int J Lab Hematol 39(3):261–271

    Article  CAS  PubMed  Google Scholar 

  28. Donohue RE, Marcogliese AN, Sasa GS, Elghetany MT, Redkar AA, Bertuch AA, Curry CV (2018) Standardized high-sensitivity flow cytometry testing for paroxysmal nocturnal hemoglobinuria in children with acquired bone marrow failure disorders: a single center us study. Cytom Part B 94:699–704

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors are thankful to Sir Ganga Ram Hospital for providing infrastructure to conduct the study.

Author information

Authors and Affiliations

  1. Department of Clinical Hematology, Sir Ganga Ram Hospital, New Delhi, India

    Faran Naim, Nitin Gupta & Ajay Sharma

  2. Department of Hematology, Sir Ganga Ram Hospital, New Delhi, 110060, India

    Amrita Saraf, Jasmita Dass, Vandana Arya, Sabina Langer & Jyoti Kotwal

Authors
  1. Faran Naim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amrita Saraf
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jasmita Dass
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vandana Arya
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sabina Langer
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Nitin Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ajay Sharma
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jyoti Kotwal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Amrita Saraf.

Ethics declarations

Conflict of interest

The authors have declared no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Naim, F., Saraf, A., Dass, J. et al. A Prospective, Cross Sectional Study of PNH Clone in MDS Patients Using High Sensitivity Flowcytometry: A Single Center Experience. Indian J Hematol Blood Transfus 36, 519–525 (2020). https://doi.org/10.1007/s12288-019-01219-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12288-019-01219-1

Keywords

Search

Navigation