title: Comparative analysis between RQ-PCR and digital-droplet-PCR of immunoglobulin/T-cell receptor gene rearrangements to monitor minimal residual disease in acute lymphoblastic leukaemia.
abstract: Real-time quantitative polymerase chain reaction (RQ-PCR) is a standardized tool for minimal residual disease (MRD) monitoring in acute lymphoblastic leukaemia (ALL). The applicability of this technology is limited by the need of a standard curve based on diagnostic DNA. The digital droplet PCR (ddPCR) technology has been recently applied to various medical fields, but its use in MRD monitoring is under investigation. In this study, we analysed 50 ALL cases by both methods in two phases: in the first, we established analytical parameters to investigate the applicability of this new technique; in the second, we analysed MRD levels in 141 follow-up (FU) samples to investigate the possible use of ddPCR for MRD monitoring in ALL patients. We documented that ddPCR has sensitivity and accuracy at least comparable to those of RQ-PCR. Overall, the two methods gave concordant results in 124 of the 141 analysed MRD samples (88%, P = 0·94). Discordant results were found in 12% borderline cases. The results obtained prove that ddPCR is a reliable method for MRD monitoring in ALL, with the advantage of quantifying without the need of the calibration curves. Its application in a cohort of patients with a longer FU will conclusively define its clinical predictive value.
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Della Starza
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Vittorio
authors.initials: V
authors.lastname: Nunes
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Marzia
authors.initials: M
authors.lastname: Cavalli
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Lucia Anna
authors.initials: LA
authors.lastname: De Novi
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Ilari
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Valerio
authors.initials: V
authors.lastname: Apicella
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Vitale
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Testi
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Del Giudice
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Sabina
authors.initials: S
authors.lastname: Chiaretti
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Robin
authors.initials: R
authors.lastname: Foà
authors.affiliation: Department of Cellular Biotechnologies and Haematology, "Sapienza" University of Rome, Rome, Italy. guarini@bce.uniroma1.it.
authors.forename: Anna
authors.initials: A
authors.lastname: Guarini
mesh_terms: D000293:Adolescent; D000328:Adult; D019411:Clinical Laboratory Techniques; D015321:Gene Rearrangement; D005803:Genes, Immunoglobulin; D019672:Genes, T-Cell Receptor; D006801:Humans; D008722:Methods; D018365:Neoplasm, Residual / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000473:pathology*; D012680:Sensitivity and Specificity; D055815:Young Adult
keywords: ALL; MRD; gene rearrangement; quantitative PCR