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

Authors :
Law SC
Haigh OL
Walpole CM
Keane C
Miles JJ
Gandhi MK
Radford KJ
Steptoe RJ
Source :
Journal of immunological methods [J Immunol Methods] 2019 Feb; Vol. 465, pp. 72-76. Date of Electronic Publication: 2018 Dec 08.
Publication Year :
2019

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.<br /> (Copyright © 2018 Elsevier B.V. All rights reserved.)

Details

Language :
English
ISSN :
1872-7905
Volume :
465
Database :
MEDLINE
Journal :
Journal of immunological methods
Publication Type :
Academic Journal
Accession number :
30537479
Full Text :
https://doi.org/10.1016/j.jim.2018.12.002