Sequence-based typing of HLA class II DQB1

Due to the expanding number of known HLA class II DQB1 alleles, high-resolu tion oligotyping is becoming ineffective, therefore a sequence-based typing (SBT) strategy was developed to provide rapid and definitive typing of HLA -DQB1. HLA-DQB1*02, *03, *04, *05, and *06 alleles were individually amplif ied by polymerase chain reaction (PCR) using tron 2 group-specific primers. Forward and reverse PCR primers were tailed with M13 universal and M13 rev erse sequences, respectively. Subsequent bi-directional cycle-sequencing wa s carried out using Cy5.5-labeled M13 universal primer and Cy5.0-labeled M1 3 reverse primer. Automated sequencing was performed in 30 min using a Visi ble Genetics, Inc. (VGI) MicroGene Clipper(TM) Sequencer. Full concordance was observed between this SET method per and oligotyping among 151 individu als.