Reference strand mediated conformation analysis resolves HLA‐DRB1 typing ambiguities when matching for unrelated bone marrow transplantation

Abstract

We show here the use of reference strand mediated conformation analysis (RSCA) to unambiguously resolve the HLA-DRB1 typing of two individuals which were selected as potential unrelated donors for bone marrow transplantation (BMT). In the first case, both sequence-specific primer (SSP) amplification and sequence-specific oligonucleotide probing (SSO), routinely used in different tissue typing laboratories gave, for the two unrelated donors, the same ambiguous typing of HLA-DRB1*04011+*0403 or DRB1*0407+*0413. In this case sequence-based typing (SBT) was not the method of choice to resolve the situation, due to the sequence ambiguities of these two given combinations. RSCA of both samples, using homozygous typing cells (HTCs) for DRB1*04011, *0403 and *0407 as internal controls, gave the unambiguous result that both donors were HLA-DRB1*04011+*0403. In the second case, a donor was typed as DRB1*1102+1103 by SSP, while SSO excluded the DRB1*1102 allele. The patient was unambiguously typed as DRB1*1101+1103 by both techniques. RSCA, using DNA from reference cell lines as internal controls, gave the unambiguous typing that the donor was DRB1*1103 homozygous.

Two sets of HLA Class II DRB and DQB1 alleles co‐segregate among family members in a single maternal haplotype

“…It is possible that the multiplicity of HLA types in this family is a common feature among maternal cousins of the patient. The risk of severe GVHD following bone marrow transplantation is significantly increased when patient and donor are HLA mismatched at more than one locus (8). A HLA matched unrelated bone marrow donor for any individual carrying three DRB1 and three DQB1 genes would be virtually impossible to find, as all donors would be both DR and DQ mismatched.…”

RSCA genotyping of MHC for high-throughput evolutionary studies in the model organism three-spined stickleback Gasterosteus aculeatus

“…This mechanism provides a significant advantage over other commonly used indirect typing techniques because it produces as many mobility values (bands or peaks) per allele as desired by using several distinct reference strands, creating a multi-dimensional coordinate for each allele. This enables differentiation of highly similar sequence variants [ 23 ].…”

Cost-effectiveness analysis of two strategies for typing unrelated donors for bone marrow transplantation in France

“…Collaboration with nonmedical disciplines for designing robust data bases, convenient methods for initiating and monitoring donor searches, and better communication between centers can also be envisaged [3,4,5]. Improvements in laboratory techniques have also been considered [6,7,8]. Generally speaking, economic pressure has emerged as an important factor.…”