Faculty Collaboration Database

Sequence-based typing of HLA class II DQB1. Tissue Antigens 2000 Apr;55(4):364-8

Date

06/14/2000

Pubmed ID

10852389

DOI

10.1034/j.1399-0039.2000.550411.x

Scopus ID

2-s2.0-0034013698 (requires institutional sign-in at Scopus site)   11 Citations

Abstract

Due to the expanding number of known HLA class II DQB1 alleles, high-resolution 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 amplified by polymerase chain reaction (PCR) using exon 2 group-specific primers. Forward and reverse PCR primers were tailed with M13 universal and M13 reverse sequences, respectively. Subsequent bi-directional cycle-sequencing was carried out using Cy5.5-labeled M13 universal primer and Cy5.0-labeled M13 reverse primer. Automated sequencing was performed in 30 min using a Visible Genetics, Inc. (VGI) MicroGene Clipper Sequencer. Full concordance was observed between this SBT method and oligotyping among 151 individuals.

Author List

Dinauer DM, Luhm RA, Uzgiris AJ, Eckels DD, Hessner MJ

Author

Martin J. Hessner PhD Professor in the Pediatrics department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

DNA Primers
Exons
Genotype
HLA-DQ Antigens
HLA-DQ beta-Chains
Histocompatibility Testing
Humans
Polymerase Chain Reaction
Sequence Analysis, DNA