47-P

Abstract

Aim Every year, thousands of people of all ages are diagnosed with leukemia and other life threatening diseases. Many of them will die unless they get a bone marrow or cord blood transplant from a matching donor. Seventy percent of people do not have a donor in their family. For those patients, donors are recruited from international registries such as the National Donor Program (NMDP) to find an unrelated matching donor. Donor registries have been set up to acquire a large set of potential donors which have been HLA genotyped. Donors are recruited based on matching criteria for donor and patient HLA types. To be able to use the registries for donor searches, the typing data should fulfill minimum requirements. For newly recruited potential donors, the NMDP has set the requirements: Donors should be typed to at least intermediate resolution, uniquely identifying the gene region that codes for the peptide binding groove, of HLA-A,-B and -DRB1 typing for cord-blood units and additionally HLA-C and -DQB1 are required for bone marrow transplantation. To efficiently type large numbers of samples, easy and quick DNA-based methods for intermediate resolution typing are required. Methods We initiated development of Sequencing-based Typing High-Throughput (SBT-HT) methods for identification of HLA-A, -B, -C, -DQB1 and DRB1 alleles at the intermediate resolution level. After amplification an adjusted Sequencing Based Typing (SBT) strategy is exploited that provides high resolution typing results, in most cases even in a single round. Results Here we show that intermediate typing resolution is obtained within one working day by analyzing exon 2 and 3 for class I and exon 2 for class II using generic and group-specific sequencing primers. Conclusions This high-throughput SBT method provides intermediate resolution typing for the relevant alleles within one working day. Westerink: GenDx: Employee. Adema: GenDx: Employee. Mulder: GenDx: Employee; Stockholder. Rozemuller: GenDx: Employee; Stockholder.