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Whole‐genome amplification for the detection of molecular targets and minimal residual disease monitoring in acute lymphoblastic leukaemia
Author(s) -
Della Starza Irene,
De Novi Lucia Anna,
Nunes Vittorio,
Del Giudice Ilaria,
Ilari Caterina,
Marinelli Marilisa,
Negulici Alina Delia,
Vitale Antonella,
Chiaretti Sabina,
Foà Robin,
Guarini Anna
Publication year - 2014
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/bjh.12744
Subject(s) - minimal residual disease , genomic dna , polymerase chain reaction , dna , real time polymerase chain reaction , biology , digital polymerase chain reaction , multiple displacement amplification , microbiology and biotechnology , genome , dna extraction , genetics , gene , leukemia
Summary Accurate genomic characterization requires sufficient amounts of optimal quality DNA . An approach for increasing the DNA amount is the whole‐genome amplification ( WGA ) method. We applied WGA to the molecular quantification and minimal residual disease ( MRD ) evaluation of acute lymphoblastic leukaemia ( ALL ), aiming to compare the results obtained from genomic DNA and amplified DNA with WGA , and to evaluate the applicability and the reliability of WGA ‐ DNA . Twenty paired samples from adult ALL patients were sequenced to identify the functional germline V ‐ D ‐ J segment at diagnosis; real‐time quantitative polymerase chain reaction ( RQ ‐ PCR ) quantitative analysis was performed both at diagnosis and follow‐up. Genomic DNA and WGA ‐ DNA screening identified equivalent 87 rearrangements. At diagnosis, the quantitative evaluation of genomic DNA samples showed 1 logarithm difference to WGA ‐ DNA samples; these levels are comparable, being within the degree of acceptability and confidence. In the follow‐up samples, RQ ‐ PCR analysis on genomic DNA and WGA showed concordant MRD results in 16/18 samples, while 2/18 were MRD ‐positive outside the quantitative range by RQ ‐ PCR (i.e. <5 × 10 −5 ) on genomic DNA and MRD ‐negative on WGA ‐ DNA . WGA ‐ DNA enables: (i) the design of accurate targets for MRD evaluation in ALL patients, (ii) accurate disease quantification at diagnosis, (iii) MRD quantification comparable to genomic DNA .