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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
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Dr Julie Irving
Irving JAE, Jesson J, Virgo P, Case MC, Minto CLJ, Eyre L, Noel N, Johansson U, Macey M, Knotts L, Helliwell M, Davies P, Whitby L, Barnett D, Hancock J, Goulden N, Lawson S
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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
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