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ISHAGE protocol: are we doing it correctly?

Alison Whitby1, Liam Whitby, Matthew Fletcher

  • 1UK NEQAS for Leucocyte Immunophenotyping, Department of Haematology, Royal Hallamshire Hospital, Sheffield S10 2QN.

Cytometry. Part B, Clinical Cytometry
|September 15, 2011
PubMed
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Incorrect use of the ISHAGE protocol for CD34(+) stem cell enumeration is common, doubling failure rates in quality assessments. Proper ISHAGE protocol implementation is crucial for accurate stem cell collections and engraftment capability evaluations.

Area of Science:

  • Hematology
  • Clinical Laboratory Science
  • Flow Cytometry

Background:

  • CD34(+) stem cell enumeration is vital for optimizing stem cell collections and assessing engraftment potential.
  • The International Society for Hematology and Blood Transfusion (ISHAGE) protocol is the most common method for flow cytometric CD34(+) stem cell enumeration.
  • Previous findings identified laboratories with persistent poor performance due to incorrect ISHAGE protocol usage.

Purpose of the Study:

  • To investigate the extent of incorrect ISHAGE protocol usage among participants in a CD34(+) stem cell enumeration External Quality Assessment (EQA) program.
  • To determine the impact of incorrect ISHAGE protocol implementation on EQA performance.

Main Methods:

  • A survey was distributed to 255 participants of the UK NEQAS CD34(+) stem cell enumeration program.

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  • Flow cytometry data and dot plots were collected and analyzed.
  • Participants' adherence to and correct setup of the ISHAGE protocol gating strategy were assessed.

    • Main Results:

    • Out of 103 laboratories providing dot plots, 83 reported using the ISHAGE protocol, but 43% (36/83) had incorrect setups.
    • Laboratories incorrectly using the single-platform ISHAGE gating strategy were twice as likely to fail an EQA exercise.
    • Incorrect ISHAGE protocol use in a dual-platform setting doubled the failure rate.

    Conclusions:

    • A significant lack of understanding and incorrect application of the ISHAGE protocol is prevalent.
    • Improper ISHAGE protocol usage has substantial implications for the accuracy of stem cell enumeration and clinical decisions.
    • Rigorous monitoring and education are necessary to improve participant performance in deploying the ISHAGE protocol.