Whole-genome amplification for the detection of molecular targets and minimal residual disease monitoring in acute lymphoblastic leukaemia.
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.
Haematology, Department of Cellular Biotechnologies and Haematology, "Sapienza" University, Rome, Italy.
Irene
I
Della Starza
Lucia Anna
LA
De Novi
Vittorio
V
Nunes
Ilaria
I
Del Giudice
Caterina
C
Ilari
Marilisa
M
Marinelli
Alina Delia
AD
Negulici
Antonella
A
Vitale
Sabina
S
Chiaretti
Robin
R
Foà
Anna
A
Guarini
D000293:Adolescent; D000328:Adult; D005260:Female; D005803:Genes, Immunoglobulin; D015894:Genome, Human; D006801:Humans; D008297:Male; D018365:Neoplasm, Residual; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000175:diagnosis* / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D055815:Young Adult
D011948:Receptors, Antigen, T-Cell
acute lymphoblastic leukaemia; immunoglobulin gene rearrangement; minimal residual disease; quantitative PCR; whole-genome amplification