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