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.
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.
Irving
Julie
J
Northern Institute for Cancer Research, Newcastle upon Tyne, Tyne and Wear, UK. j.a.e.irving@ncl.ac.uk
Jesson
Jenny
J
Virgo
Paul
P
Case
Marian
M
Minto
Lynne
L
Eyre
Lisa
L
Noel
Nigel
N
Johansson
Ulrika
U
Macey
Marion
M
Knotts
Linda
L
Helliwell
Margaret
M
Davies
Paul
P
Whitby
Liam
L
Barnett
David
D
Hancock
Jeremy
J
Goulden
Nick
N
Lawson
Sarah
S
D018941:Antigens, CD19 / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D002648:Child; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D015321:Gene Rearrangement; D006801:Humans; D015448:Leukemia, B-Cell / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D018365:Neoplasm, Residual / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D015260:Neprilysin / Q000032:analysis; D016133:Polymerase Chain Reaction; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D011379:Prognosis; D011446:Prospective Studies; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D018941:Antigens, CD19; D018952:Antigens, CD34; D011948:Receptors, Antigen, T-Cell; D015260:Neprilysin