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Exploration of methods for sequence based HLA typing and application to patients with hair dye allergy Open Access


Other title
Human Leukocyte Antigens
Hair dye allergy
HLA typing
Type of item
Degree grantor
University of Alberta
Author or creator
Garcia-Batres, Carlos R.
Supervisor and department
Elliott, John (Medical Microbiology and Immunology)
Examining committee member and department
Magor, Katharine (Biological Sciences)
Evans, David (Medical Microbiology and Immunology)
Burshtyn, Deborah (Medical Microbiology and Immunology)
Department of Medical Microbiology and Immunology

Date accepted
Graduation date
Master of Science
Degree level
Determining an individual’s spectrum of human leukocyte antigens (i.e. HLA type) by sequencing their HLA genes is known as sequenced-based typing (SBT). In the first part of this thesis, non-polymerase chain reaction (PCR) methods of achieving SBT were explored. HLA-DQ cDNA sequences were cloned into a novel M13 vector so that antisense sequence was displayed, and attempts were made to specifically capture the DQ sequences using two strategies: 1) complementary oligonucleotides covalently bound to the surface of beads, and 2) complementary biotinylated oligonucleotides which were then captured using streptavadin-beads. Only the second protocol was successful, enabling enrichment of sequences about 100-fold, but the method was deemed unsuitable to achieve SBT. In the second part of this thesis an RT-PCR method to amplify the entire coding region of HLA-DPB1 transcripts was developed. By cloning and sequencing the amplified fragments, the HLA-DPB1 type could be assigned. Methods were also developed to extract total cellular RNA from human blood samples, and RNA samples from 16 hair dye-allergic patients were obtained. These were subjected to RT-PCR and HLA-DPB1 alleles assigned. The allelic distribution of the patient samples did not differ from a control population, suggesting that hair dye allergy is not associated with certain HLA-DPB1 alleles.
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
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