title: Next-generation sequencing indicates false-positive MRD results and better predicts prognosis after SCT in patients with childhood ALL.
abstract: Minimal residual disease (MRD) monitoring via quantitative PCR (qPCR) detection of Ag receptor gene rearrangements has been the most sensitive method for predicting prognosis and making post-transplant treatment decisions for patients with ALL. Despite the broad clinical usefulness and standardization of this method, we and others have repeatedly reported the possibility of false-positive MRD results caused by massive B-lymphocyte regeneration after stem cell transplantation (SCT). Next-generation sequencing (NGS) enables precise and sensitive detection of multiple Ag receptor rearrangements, thus providing a more specific readout compared to qPCR. We investigated two cohorts of children with ALL who underwent SCT (30 patients and 228 samples). The first cohort consisted of 17 patients who remained in long-term CR after SCT despite having low MRD positivity (<0.01%) at least once during post-SCT monitoring using qPCR. Only one of 27 qPCR-positive samples was confirmed to be positive by NGS. Conversely, 10 of 15 samples with low qPCR-detected MRD positivity from 13 patients who subsequently relapsed were also confirmed to be positive by NGS (P=0.002). These data show that NGS has a better specificity in post-SCT ALL management and indicate that treatment interventions aimed at reverting impending relapse should not be based on qPCR only.
authors.affiliation: CLIP Laboratory Centre, Department of Pediatric Hematology/Oncology, 2nd Faculty of Medicine and University Hospital Motol, Prague, Czech Republic.
authors.forename: M
authors.initials: M
authors.lastname: Kotrova
authors.affiliation: Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands.
authors.forename: V H J
authors.initials: VHJ
authors.lastname: van der Velden
authors.affiliation: Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands.|Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: J J M
authors.initials: JJM
authors.lastname: van Dongen
authors.affiliation: Department of Pediatric Hematology/Oncology, 2nd Faculty of Medicine and University Hospital Motol, Prague, Czech Republic.
authors.forename: R
authors.initials: R
authors.lastname: Formankova
authors.affiliation: Department of Pediatric Hematology/Oncology, 2nd Faculty of Medicine and University Hospital Motol, Prague, Czech Republic.
authors.forename: P
authors.initials: P
authors.lastname: Sedlacek
authors.affiliation: Department of Hematology, University Hospital Schleswig-Holstein, Kiel, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Brüggemann
authors.affiliation: CLIP Laboratory Centre, Department of Pediatric Hematology/Oncology, 2nd Faculty of Medicine and University Hospital Motol, Prague, Czech Republic.
authors.forename: J
authors.initials: J
authors.lastname: Zuna
authors.affiliation: Department of Pediatric Hematology/Oncology, 2nd Faculty of Medicine and University Hospital Motol, Prague, Czech Republic.
authors.forename: J
authors.initials: J
authors.lastname: Stary
authors.affiliation: CLIP Laboratory Centre, Department of Pediatric Hematology/Oncology, 2nd Faculty of Medicine and University Hospital Motol, Prague, Czech Republic.
authors.forename: J
authors.initials: J
authors.lastname: Trka
authors.affiliation: CLIP Laboratory Centre, Department of Pediatric Hematology/Oncology, 2nd Faculty of Medicine and University Hospital Motol, Prague, Czech Republic.
authors.forename: E
authors.initials: E
authors.lastname: Fronkova
mesh_terms: D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D005189:False Positive Reactions; D005260:Female; D018380:Hematopoietic Stem Cell Transplantation; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D018365:Neoplasm, Residual; D016133:Polymerase Chain Reaction; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000097:blood / Q000175:diagnosis / Q000235:genetics / Q000628:therapy; D011379:Prognosis