A novel method for simultaneous high resolution identification of HLA-A, HLA-B, and HLA-Cw alleles.

AUTOR(ES)

Arguello, R

RESUMO

We describe a novel high resolution DNA based typing approach for HLA class I alleles, which identifies the recombinational motifs present in exons 2 and 3 of the HLA class I genes. Unique identification patterns for 201 known HLA-A, HLA-B, and HLA-Cw alleles were generated by the use of only 40 probes, which were targeted at these common motifs. The unambiguous identification of the alleles was achieved by the development of a new and powerful allelic separation technique that allows isolation of single alleles after amplification. To validate the method, we have used locus-specific primers to amplify exons 2 and 3 of HLA-A, HLA-B, and HLA-Cw loci from 22 heterozygous and 41 homozygous cell lines. After amplification, the allelic fragments from each locus were separated, blotted, and hybridized with the 40 probes. In all cases, the allelic products could be separated and 81 different class I alleles, 33 HLA-A, 30 HLA-B, and 18 HLA-Cw, were identified according to the predicted probe hybridization patterns.

Documentos Relacionados

• HLA-Cw allele analysis by PCR-restriction fragment length polymorphism: study of known and additional alleles.
• Performance Characteristics of Updated INNO-LiPA Assays for Molecular Typing of Human Leukocyte Antigen A (HLA-A), HLA-B, and HLA-DQB1 Alleles
• Nature of polymorphism in HLA-A, -B, and -C molecules.
• Complete sequence of HLA-B27 cDNA identified through the characterization of structural markers unique to the HLA-A, -B, and -C allelic series.
• Associação do HLA-B*14 e HLA-Cw*08 com a suscetibilidade para vasculite reumatóide (VR) e HLA-DRB5*01 na proteção para VR em pacientes brasileiros