title: HLA-B locus sequence-based typing.
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.
authors: National Cancer Institute, IST, c/o Advanced Biotechnology Center, Genoa, Italy.
authors: S
authors: S
authors: Pozzi
authors: A
authors: A
authors: Longo
authors: G B
authors: GB
authors: Ferrara
mesh_terms: D000483:Alleles; D002874:Chromosome Mapping; D005838:Genotype; D015235:HLA-B Antigens / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D015235:HLA-B Antigens