Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Technical Report
  • Published:

Assessing agreement between CD34 enumeration by flow cytometry and volumetric analysis

Abstract

Prior to replacement of an established method for CD34 enumeration by an alternative approach, evaluation of the agreement between the methods is essential. In this study, the comparison of two assays was evaluated according to the recommendation of Bland and Altman describing the agreement between two methods where the true value is not known. CD34 enumeration was performed on blood or leukapheresis product from 105 patients by flow cytometry (dual platform assay) and volumetric analysis (single platform assay). Both the flow cytometric and the volumetric analysis showed poor reproducibility for measures lower than approximately 9 CD34+ cells/mm3. For values higher than 29 CD34+ cells/mm3, evaluation of the agreement demonstrated a difference between the single and dual platform assay, where CD34 enumeration by the volumetric analysis demonstrated values 73–80% of the flow cytometric value. The difference between the two assays could be due to several technical pitfalls which are discussed.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  1. Serke S, Johnsen HE . A European reference protocol for quality assessment and clinical validation of autologous haematopoietic blood progenitor and stem cell grafts Bone Marrow Transplant 2001 27: 463 470

    Article  CAS  Google Scholar 

  2. Wunder E, Sovalat H, Fritsch G et al. Report on the European Workshop on Peripheral Blood Stem Cell Determination and Standardization – Mulhouse, France, February 6–8 and 14–15. 1992 J Hematother 1992 1: 131 142

    Article  CAS  Google Scholar 

  3. Sutherland DR, Anderson L, Keeney M et al. The ISHAGE guidelines for CD34+ cell determination by flow cytometry. International Society of Hematotherapy and Graft Engineering J Hematother 1996 5: 213 226

    Article  CAS  Google Scholar 

  4. Serke S, Sauberlich S, Huhn D . Multiparameter flow-cytometrical quantitation of circulating CD34(+)-cells: correlation to the quantitation of circulating haemopoietic progenitor cells by in vitro colony-assay Br J Haematol 1991 77: 453 459

    Article  CAS  Google Scholar 

  5. Johnsen HE, Baech J, Nicolaisen K et al. Validation of the Nordic Flow Cytometry Standard for CD34+ Cell Enumeration in Blood and Autografts: Report from the Third Workshop J Hematother 1999 8: 15 28

    Article  CAS  Google Scholar 

  6. Bland JM, Altman DG . Statistical methods for assessing agreement between two methods of clinical measurement Lancet 1986 1: 307 310

    Article  CAS  Google Scholar 

  7. Knudsen LM, Gaarsdal E, Jensen L et al. Evaluation of mobilized CD34+ cell counts to guide timing and yield of large-scale collection by leukapheresis J Hematother 1998 7: 45 52

    Article  CAS  Google Scholar 

  8. Bland JM, Altman DG . Measuring agreement in method comparison studies Stat Meth Med Res 1999 8: 135 160

    Article  CAS  Google Scholar 

  9. Read EJ, Kunitake ST, Carter CS et al. Enumeration of CD34+ hematopoietic progenitor cells in peripheral blood and leukapheresis products by microvolume fluorimetry: a comparison with flow cytometry J Hematother 1997 6: 291 301

    Article  CAS  Google Scholar 

  10. Cabezudo E, Querol S, Cancelas JA et al. Comparison of volumetric capillary cytometry with standard flow cytometry for routine enumeration of CD34+ cells Transfusion 1999 39: 864 872

    Article  CAS  Google Scholar 

  11. Gratama JW, Orfao A, Barnett D et al. Flow cytometric enumeration of CD34+ hematopoietic stem and progenitor cells. European Working Group on Clinical Cell Analysis Cytometry 1998 34: 128 142

    Article  CAS  Google Scholar 

  12. Keeney M, Chin-Yee I, Weir K et al. Single platform flow cytometric absolute CD34+ cell counts based on the ISHAGE guidelines. International Society of Hematotherapy and Graft Engineering (see comments) Cytometry 1998 34: 61 70

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was in part aided by grants from The Danish Blood Donors Research Foundation.

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gisselø, C., Roer, O., Hoffmann, M. et al. Assessing agreement between CD34 enumeration by flow cytometry and volumetric analysis. Bone Marrow Transplant 29, 699–703 (2002). https://doi.org/10.1038/sj.bmt.1703514

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.bmt.1703514

Keywords

Search

Quick links