Simple, rapid and inexpensive typing of common HLA class I alleles for immunological studies

Law, Soi Cheng, Haigh, Oscar L., Walpole, Carina M., Colm, Keane, Miles, John J., Ghandi, Maher K., Radford, Kristen J., and Steptoe, Raymond J. (2019) Simple, rapid and inexpensive typing of common HLA class I alleles for immunological studies. Journal of Immunological Methods, 465. pp. 72-76.

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Abstract

Current HLA-typing methods are typically designed to provide exquisitely-detailed identification of multiple HLA-alleles to satisfy the requirements for organ and bone marrow transplantation or genetic studies. Many human immunological studies, on the other hand, focus around only a small number of HLA alleles that are abundant or of relevance to specific diseases. Consequently, for such studies, many HLA typing approaches are not cost-effective and are potentially complicated, slow and not easily performed in-house. Work-flow would be streamlined by a simple, inexpensive and rapid typing method able to be performed in-house. We outline a straightforward approach that provides appropriate data for much immunological research. In a predominantly Caucasian population, flow cytometry using anti-HLA-A2, -B8 and -B7 antibodies consistently and accurately screened for samples carrying the highly-abundant HLA class I alleles HLA-A*02:01, -B*08:01 and -B*07:02 that form the focus of immunological studies. Next, we describe a straightforward and simple strategy for design and use of allele-specific PCR primers to identify, at high-resolution, alleles of interest. When combined with a simple gDNA extraction technique this provides reliable, simple and inexpensive in-house HLA typing demonstrated here for highly-abundant HLA class I alleles.

Item ID: 57195
Item Type: Article (Research - C1)
ISSN: 1872-7905
Keywords: HLA typing; PCR; flow cytometry
Copyright Information: Copyright © 2018 Elsevier B.V. All rights reserved.
Funders: Cancer Council Queensland (CCQ), University of Queensland (UQ), Mater Foundation, US Department of Defense (DOD), Leukemia Foundation
Projects and Grants: CCQ 1098690, UQ Vice Chancellor's Senior Research Fellowship, Mater Foundation Senior Research Fellowship, DOD grant W81XWH-15-0451
Date Deposited: 26 Feb 2019 22:19
FoR Codes: 11 MEDICAL AND HEALTH SCIENCES > 1107 Immunology > 110799 Immunology not elsewhere classified @ 100%
SEO Codes: 92 HEALTH > 9201 Clinical Health (Organs, Diseases and Abnormal Conditions) > 920199 Clinical Health (Organs, Diseases and Abnormal Conditions) not elsewhere classified @ 100%
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