Concurrent detection of minimal residual disease (MRD) in childhood acute lymphoblastic leukaemia by flow cytometry and real-time PCR.
Minimal (i.e. submicroscopic) residual disease (MRD) predicts outcome in childhood acute lymphoblastic leukaemia (ALL). To be used clinically, MRD assays must be reliable and accurate. Two well-established techniques, flow cytometry (FC) and polymerase chain reaction (PCR), can detect leukaemic cells with a sensitivity of 0.01% (10(-4)). We analysed diagnostic samples of 45 ALL-patients (37 B-lineage ALL, eight T-lineage ALL) by four-colour FC and real-time PCR. Leukaemia-associated immunophenotypes, at a sensitivity of MRD detection by FC at the 0.01% level, were identified in 41 cases (91%); antigen-receptor gene rearrangements suitable for MRD detection with a sensitivity of 0.01% or better by PCR were identified in 38 cases (84%). The combined use of FC and PCR allowed MRD monitoring in all 45 patients. In 105 follow-up samples, MRD estimates by both methods were highly concordant, with a deviation factor of <5 by Bland-Altman analysis. Importantly, the concordance between FC and PCR was also observed in regenerating bone marrow samples containing high proportions of CD19(+) cells, and in samples studied 24 h after collection. We conclude that both MRD assays yield generally concordant results. Their combined use should enable MRD monitoring in virtually all patients and prevent false-negative results due to clonal evolution or phenotypic shifts.
Kerst
Gunter
G
Department of Pediatric Haematology and Oncology, University Children's Hospital, D-72076 Tübingen, Germany. gunter.kerst@med.uni-tuebingen.de
Kreyenberg
Hermann
H
Roth
Carmen
C
Well
Catrin
C
Dietz
Klaus
K
Coustan-Smith
Elaine
E
Campana
Dario
D
Koscielniak
Ewa
E
Niemeyer
Charlotte
C
Schlegel
Paul G
PG
Müller
Ingo
I
Niethammer
Dietrich
D
Bader
Peter
P
D000293:Adolescent; D014408:Biomarkers, Tumor / Q000032:analysis*; D002648:Child; D002675:Child, Preschool; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D015329:Gene Rearrangement, T-Lymphocyte; D006801:Humans; D007223:Infant; D018365:Neoplasm, Residual; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000473:pathology*; D012680:Sensitivity and Specificity
D014408:Biomarkers, Tumor