Journal Article

Earlier relapse detection after allogeneic haematopoietic stem cell transplantation by chimerism assays: Digital PCR versus quantitative real-time PCR of insertion/deletion polymorphisms

Valero-Garcia, Jennifer, González-Espinosa, María del Carmen, García-Planells, Javier, Ruiz-Lafora, Carlos, Barrios, Manuel, Fuentes-Gálvez, Ainhoa, Carmona-Antoñanzas, Greta, Jiménez-Velasco, Antonio

2019

Overview

The analysis of molecular haematopoietic chimerisms (HC) has become a well-established method to monitor the transplant evolution and to assess the risk of relapse after allogeneic stem cells transplantation (allo-STC). Different techniques and molecular markers are being used for chimerism surveillance after transplantation, including quantitative real-time PCR (qPCR) and the recently developed digital PCR (dPCR). This study aims to compare the sensitivity and accuracy of both methods to quantify HC and predict early relapse. HC was evaluated using custom PCR systems for the specific detection of the Y-chromosome, null alleles and insertion-deletion polymorphisms. A total of 281 samples from 28 adult patients who underwent an allo-SCT were studied. Increasing mixed chimerism was detected prior to relapse in 100% of patients (18 relapses). Compared with conventional qPCR amplification, dPCR predicted relapse with a median anticipation period of 63 days versus 45.5 days by qPCR. Overall, 56% of the relapses were predicted earlier with dPCR whereas 38% of the relapses where detected simultaneously using both techniques and only in 1 case, relapse was predicted earlier with qPCR. In conclusion, chimerism determination by dPCR constitutes a suitable technique for the follow-up of patients with haematological pathologies after allo-STC, showing greater sensitivity to predict an early relapse.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Accuracy / Adult / Alleles / Analysis / Biology and Life Sciences / Bone marrow / Chimerism / Chromosome deletion / Clonal deletion / Disease / EDTA / Female / Genetic aspects / Hematologic Neoplasms - genetics / Hematologic Neoplasms - therapy / Hematology / Hematopoietic Stem Cell Transplantation / Hematopoietic stem cells / Hospitals / Humans / INDEL Mutation / Insertion / Intelligence gathering / Leukemia / Male / Medical research / Medicine and Health Sciences / Methods / Middle Aged / Monitoring methods / Mutation / Patients / Polymerase chain reaction / Polymorphism, Genetic / Real time / Real-Time Polymerase Chain Reaction / Recurrence / Recurrence (Disease) / Research and Analysis Methods / Risk assessment / Sensitivity / Stem cell transplantation / Stem cells / Surveillance / Transplantation / Transplantation Chimera - genetics / Transplantation, Homologous / Transplants & implants / Y chromosomes