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PubMed · 10203021

HLA-B locus sequence-based typing.

S Pozzi·A Longo·G B Ferrara
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Abstract

We describe an approach for HLA-B high-resolution typing. A single locus-specific amplification generates a 1-kb fragment useful for direct sequencing. Four internal primers are necessary for exon 2 and 3 cycle-sequencing in both directions. Fluorescent dye-labelled nucleotides are incorporated during cycle-sequencing and reaction products are analyzed in an automated DNA sequencer. At present, software programs allow automatic assignment of exon 2 only; analysis of exon 3 is not automatic. In the future, the development of more sophisticated software will improve allele assignment. The approach described in this work offers a precise and efficient identification of known allele sequences and at the same time can differentiate new alleles. Furthermore, it may be applied as a model for the development of similar molecular typing approaches for other polymorphic HLA loci.

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AllelesChromosome MappingGenotypeHLA-B AntigensHumansPolymerase Chain ReactionSequence Analysis, DNAJournal ArticleResearch Support, Non-U.S. Gov't

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BibTeXRIS

S Pozzi, A Longo, G B Ferrara. 1999. HLA-B locus sequence-based typing.. https://doi.org/10.1034/j.1399-0039.1999.530308.x

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