Published online 18 April 2009
Haematologica, Vol 94, Issue 6, 870-874 doi:10.3324/haematol.2008.000414
Copyright © 2009 by Ferrata Storti Foundation
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Brief Reports

Establishment and validation of a standard protocol for the detection of minimal residual disease in B lineage childhood acute lymphoblastic leukemia by flow cytometry in a multi-center setting;

Julie Irving1, Jenny Jesson2, Paul Virgo3, Marian Case1, Lynne Minto1, Lisa Eyre2, Nigel Noel3, Ulrika Johansson4, Marion Macey4, Linda Knotts5, Margaret Helliwell6, Paul Davies2, Liam Whitby7, David Barnett7, Jeremy Hancock3, Nick Goulden8, Sarah Lawson2 on behalf of the UKALL Flow MRD group and UK MRD steering group

1 Northern Institute for Cancer Research, Newcastle upon Tyne
2 Birmingham Children’s Hospital, Birmingham
3 Southmead Hospital, Bristol
4 The Royal London Hospital, London
5 Yorkhill Children’s Hospital, Glasgow
6 Sheffield Children’s Hospital, Sheffield
7 UK NEQAS for Leucocyte Immunophenotyping, Royal Hallamshire Hospital, Sheffield
8 Great Ormond Street Children’s Hospital, London, UK

Correspondence: Julie Irving, Northern Institute for Cancer Research, Paul O’Gorman Building, Framlington Place, Newcastle upon Tyne, Tyne and Wear, UK, NE2 4HH. E-mail.j.a.e.irving{at}ncl.ac.uk

ABSTRACT

Minimal residual disease detection, used for clinical management of children with acute lymphoblastic leukemia, can be performed by molecular analysis of antigen-receptor gene rearrangements or by flow cytometric analysis of aberrant immunophenotypes. For flow minimal residual disease to be incorporated into larger national and international trials, a quality assured, standardized method is needed which can be performed in a multi-center setting. We report a four color, flow cytometric protocol established and validated by the UK acute lymphoblastic leukemia Flow minimal residual disease group. Quality assurance testing gave high inter-laboratory agreement with no values differing from a median consensus value by more than one point on a logarithmic scale. Prospective screening of B-ALL patients (n=206) showed the method was applicable to 88.3% of patients. The minimal residual disease in bone marrow aspirates was quantified and compared to molecular data. The combined risk category concordance (minimal residual disease levels above or below 0.01%) was 86% (n=134). Thus, this standardized protocol is highly reproducible between laboratories, sensitive, applicable, and shows good concordance with molecular-based analysis.

Key words: childhood acute lymphoblastic leukemia, minimal residual disease, flow cytometry.


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