Development and performance of a next generation sequencing (NGS) assay for monitoring of mixed chimerism

Chimerism Monitoring Techniques after Hematopoietic Stem Cell Transplantation: An Overview of the Last 15 Years of Innovations

“…However, Vives et al [ 47 ] proved that STR and Indel markers can also be analyzed with these new high-throughput sequencing technologies to detect chimerism. In accordance with Pettersson et al [ 10 ], when SNP or Indel panels are used, NGS cannot be used as well in patients who were transplanted with two donors.…”

“…The most recent NGS technology can use SNP, STR, or Indel markers according to the chosen panel, resulting in a different informative rate. Moreover, the ability to work in multiplex reduces the drawbacks which can be attributed to the bi-allelic markers in this case as well [ 10 , 45 , 47 , 60 ].…”

“…NGS can only be used as a supplement for chimerism monitoring because it lacks shared experimental protocols, and it is quite complex, time-consuming, and expensive. However, despite these problems, it seems to be a very promising technique due to its high-throughput sequencing power to detect chimerism [ 10 , 31 , 33 , 60 , 61 ].…”

“…Currently, allogeneic HSCT is an efficient curative approach for different hematologic diseases, but it can also be characterized by various complications, such as toxicity related to treatment, infections, recurrence of the disease, and immunological reactions including rejection of the transplantation by the recipient and graft-versus-host disease (GVHD) [ 8 , 9 , 10 ]. The success of allogeneic HSCT is evaluated by chimerism analysis, through which the monitoring of the relative amount of living donor cells and residual recipient cells is performed in samples of peripheral blood or bone marrow; what is clinically useful is not the single determination, but the monitoring over time of the dynamics or the changes in the percentages of donor and recipient cells between different time-points [ 11 ].…”

Performance Assessment of the Devyser High-Throughput Sequencing–Based Assay for Chimerism Monitoring in Patients after Allogeneic Hematopoietic Stem Cell Transplantation

“…17 In recent years, assays using high-throughput sequencing (HTS) for HC monitoring have been developed. 16,18,19 These assays allow for HC monitoring using a single, highly multiplexed PCR reaction while showing promise in attaining an LoD and limit of quantification down to 0.1% 16,19 ; this approach in effect combines the advantages of the PCR-CEe and qPCR-based assays. The current article evaluates the Devyser NGS chimerism assay, which is a commercial HTS-based assay targeting 24 loci with a known biallelic insertion/deletion polymorphism.…”