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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
1 Northern Institute for Cancer Research, Newcastle upon Tyne
2 Birmingham Childrens Hospital, Birmingham
3 Southmead Hospital, Bristol
4 The Royal London Hospital, London
5 Yorkhill Childrens Hospital, Glasgow
6 Sheffield Childrens Hospital, Sheffield
7 UK NEQAS for Leucocyte Immunophenotyping, Royal Hallamshire Hospital, Sheffield
8 Great Ormond Street Childrens Hospital, London, UK
Correspondence: Julie Irving, Northern Institute for Cancer Research, Paul OGorman 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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