TY - JOUR
T1 - Immunoglobulin kappa deleting element rearrangements in precursor-B acute lymphoblastic leukemia are stable targets for detection of minimal residual disease by real-time quantitative PCR
AU - van der Velden, V. H. J.
AU - Willemse, M. J.
AU - van der Schoot, C. E.
AU - Hählen, K.
AU - van Wering, E. R.
AU - van Dongen, J. J. M.
PY - 2002
Y1 - 2002
N2 - Immunoglobulin gene rearrangements are used as PCR targets for detection of minimal residual disease (MRD) in acute lymphoblastic leukemia (ALL). We Investigated the occurrence of monoclonal immunoglobulin kappa-deleting element (IGK-Kde) rearrangements by Southern blotting and PCR/heteroduplex analysis at diagnosis, their stability at relapse, and their applicability in real-time quantitative PCR (RQ-PCR) analysis. In 77 selected children with precursor-B-ALL, Southern blotting detected 122 IGK-Kde rearrangements, 12 of which were derived from subclones in six patients (8%). PCR/heteroduplex analysis with BIOMED-1 Concerted Action primers identified 100 of the 110 major IGK-Kde rearrangements (91%). Comparison between diagnosis and relapse samples from 21 patients with PCR-detectable IGK-Kde rearrangements (using Southern blotting, PCR/heteroduplex analysis, and sequencing) demonstrated that 27 of the 32 rearrangements remained stable at relapse. When patients with oligoclonal IGK-Kde rearrangements were excluded, 25 of the 27 rearrangements remained stable at relapse and at least one stable rearrangement was present In 17 of the 18 patients. Subsequently, RQ-PCR analysis with allele-specific forward primers, a germline Kde Taq-Man-probe, and a germline Kde reverse primer was evaluated for 18 IGK-Kde rearrangements. In 16 of the 18 targets (89%) a sensitivity of less than or equal to10(-4) was reached. Analysis of MRD during follow-up of eight patients with IGK-Kde rearrangements showed comparable results between RQ-PCR data and classical dot-blot data. We conclude that the frequently occurring IGK-Kde rearrangements are generally detectable by PCR (similar to90%) and are highly stable MRD-PCR targets, particularly where monoclonal rearrangements at diagnosis similar to(95%) are concerned. Furthermore, most IGK-Kde rearrangements (similar to90%) can be used for sensitive detection of MRD (less than or equal to10(-4)) by RQ-PCR analysis
AB - Immunoglobulin gene rearrangements are used as PCR targets for detection of minimal residual disease (MRD) in acute lymphoblastic leukemia (ALL). We Investigated the occurrence of monoclonal immunoglobulin kappa-deleting element (IGK-Kde) rearrangements by Southern blotting and PCR/heteroduplex analysis at diagnosis, their stability at relapse, and their applicability in real-time quantitative PCR (RQ-PCR) analysis. In 77 selected children with precursor-B-ALL, Southern blotting detected 122 IGK-Kde rearrangements, 12 of which were derived from subclones in six patients (8%). PCR/heteroduplex analysis with BIOMED-1 Concerted Action primers identified 100 of the 110 major IGK-Kde rearrangements (91%). Comparison between diagnosis and relapse samples from 21 patients with PCR-detectable IGK-Kde rearrangements (using Southern blotting, PCR/heteroduplex analysis, and sequencing) demonstrated that 27 of the 32 rearrangements remained stable at relapse. When patients with oligoclonal IGK-Kde rearrangements were excluded, 25 of the 27 rearrangements remained stable at relapse and at least one stable rearrangement was present In 17 of the 18 patients. Subsequently, RQ-PCR analysis with allele-specific forward primers, a germline Kde Taq-Man-probe, and a germline Kde reverse primer was evaluated for 18 IGK-Kde rearrangements. In 16 of the 18 targets (89%) a sensitivity of less than or equal to10(-4) was reached. Analysis of MRD during follow-up of eight patients with IGK-Kde rearrangements showed comparable results between RQ-PCR data and classical dot-blot data. We conclude that the frequently occurring IGK-Kde rearrangements are generally detectable by PCR (similar to90%) and are highly stable MRD-PCR targets, particularly where monoclonal rearrangements at diagnosis similar to(95%) are concerned. Furthermore, most IGK-Kde rearrangements (similar to90%) can be used for sensitive detection of MRD (less than or equal to10(-4)) by RQ-PCR analysis
U2 - https://doi.org/10.1038/sj.leu.2402475
DO - https://doi.org/10.1038/sj.leu.2402475
M3 - Article
C2 - 11986956
SN - 0887-6924
VL - 16
SP - 928
EP - 936
JO - Leukemia
JF - Leukemia
IS - 5
ER -