A novel human leucocyte antigen-DRB1 genotyping method based on multiplex primer extension reactions

Jaakkola, E., Herzberg, I., Crane, A. M., Pointon, J. J., Laiho, K., Kauppi, M., Kaarela, K., Wordsworth, B. P., Tuomilehto, J., & Brown, M. A. (2004) A novel human leucocyte antigen-DRB1 genotyping method based on multiplex primer extension reactions. Tissue Antigens, 64(1), pp. 88-95.

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We have developed and validated a semi-automated fluorescent method of genotyping human leucocyte antigen (HLA)-DRB1 alleles, HLA-DRB1*01-16, by multiplex primer extension reactions. This method is based on the extension of a primer that anneals immediately adjacent to the single-nucleotide polymorphism with fluorescent dideoxynucleotide triphosphates (minisequencing), followed by analysis on an ABI Prism 3700 capillary electrophoresis instrument. The validity of the method was confirmed by genotyping 261 individuals using both this method and polymerase chain reaction with sequence-specific primer (PCR-SSP) or sequencing and by demonstrating Mendelian inheritance of HLA-DRB1 alleles in families. Our method provides a rapid means of performing high-throughput HLA-DRB1 genotyping using only two PCR reactions followed by four multiplex primer extension reactions and PCR-SSP for some allele groups. In this article, we describe the method and discuss its advantages and limitations.

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7 citations in Scopus
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5 citations in Web of Science®

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ID Code: 89542
Item Type: Journal Article
Refereed: Yes
Additional Information: Cited By :7
Export Date: 21 September 2015
Correspondence Address: Brown, M.A.; Inst. of Musculoskeletal Sciences, University of Oxford, Nuffield Dept. of Orthoped. Surgery, Headington, Oxford, United Kingdom; email: mbrown@well.ox.ac.uk
Keywords: High throughput, HLA typing, HLA-DRB1, Primer extension, leukocyte antigen, leukocyte antigen drb1, unclassified drug, analytic method, article, capillary electrophoresis, controlled study, fluorescent antibody technique, gene frequency, gene sequence, genotype, human, human cell, polymerase chain reaction, priority journal, sequence analysis, single nucleotide polymorphism, Alleles, Base Sequence, DNA Primers, Female, Finland, Fluorescent Dyes, HLA-DR Antigens, Humans, Male, Polymorphism, Genetic, Polymorphism, Single-Stranded Conformational, Spondylitis, Ankylosing
DOI: 10.1111/j.1399-0039.2004.00241.x
ISSN: 0001-2815
Divisions: Current > QUT Faculties and Divisions > Faculty of Health
Current > Institutes > Institute of Health and Biomedical Innovation
Copyright Owner: Copyright Blackwell Munksgaard 2004
Deposited On: 27 Oct 2015 01:25
Last Modified: 27 Oct 2015 01:25

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