Application of high‐throughput, high‐resolution and cost‐effective next generation sequencing‐based large‐scale HLA typing in donor registry

Abstract

Next generation sequencing (NGS)-based human leukocyte antigen (HLA) typing was used for ultra large-scale genotyping of registry donors for the China Marrow Donor Program (CMDP). More than 79,000 samples were subjected to HLA genotyping at 4-digit allelic level without ambiguities for HLA-A, -B, -C, DRB1 and DQB1 loci, with throughput up to 2068 samples per lane in a HiSeq flow cell (eight lanes per run), and cost reduced by 95% compared with that of Sanger-based typing. Two percent of randomly selected samples were quality control (QC) tested at 4-digit allelic level by the CMDP QC laboratory, yielded a concordance of 99.72%. These results demonstrate that NGS is a cost effective and valuable tool for HLA typing of registry donors.

Evaluation of the AllType kit for HLA typing using the Ion Torrent S5 XL platform

“…These were easily detected due to non‐common linkage disequilibrium. Most studies have reported very low allele dropout frequencies (<1%), whereas others have reported frequencies as high as 10%, especially for the HLA‐DQB1 locus . Therefore, the need for alternative methods to confirm apparently homozygous HLA alleles is a matter of debate.…”

Immunogenetics in stem cell donor registry work: The DKMS example (Part 2)

“…In this two‐part review article, we described, analysed and discussed aspects of donor registry work related to immunogenetics and population genetics by using the example of DKMS. The major development at the intersection of immunogenetics and stem cell donor registries over the last few years has clearly been the use of NGS‐based high‐throughput HLA typing (Cereb, Kim, Ryu, & Yang, ; Lange et al, ; Schöfl et al, ; Zhou et al, ). We showed that this accomplishment had important immediate effects such as higher typing quality (higher resolution, less errors) and dramatic cost savings.…”

Modified conditioning regimen improves outcomes of unrelated donor peripheral blood stem cell transplantation for β‐thalassaemia major patients

“…It is well known that patients with TM have a high rate of graft rejection due to the existence of HLA antibodies resulting from long‐term blood transfusion. The next generation sequencing HLA genotyping workshop played a potential role in selecting high compatibility of UDs in PBSCT . In our study, all patients received UD‐PBSCT with at most two allelic mismatches in addition to a high number of total nucleated cells/kg (9.6 × 10 8 /kg).…”

“…The next generation sequencing HLA genotyping workshop played a potential role in selecting high compatibility of UDs in PBSCT. 16,17 In our study, all patients received UD-PBSCT with at most two allelic mismatches in addition to a high number of total nucleated cells/kg (9.6 × 10 8 /kg).…”