HLA-Cw allele analysis by PCR-restriction fragment length polymorphism: study of known and additional alleles.
AUTOR(ES)
Tatari, Z
RESUMO
We describe a technique for HLA-Cw genotyping by digestion of PCR-amplified genes with restriction endonucleases. Locus-specific primers selectively amplified HLA-Cw sequences from exon 2 in a single PCR that avoided coamplification of other classical and nonclassical class I genes. Amplified DNAs were digested with selected enzymes. Sixty-three homozygous cell lines from International Histocompatibility Workshop X and 113 unrelated individual cells were genotypes for HLA-Cw and compared with serology. The present protocol can distinguish 23 alleles corresponding to the known HLA-Cw sequences. Genotyping of serologically undetectable alleles (HLA-Cw Blank) and of heterozygous cells was made possible by using this method. Six additional HLA-Cw alleles were identified by unusual restriction patterns and confirmed by sequencing; this observation suggests the presence of another family of allele-sharing clusters in the HLA-B locus. This PCR-restriction endonuclease method provides a simple and convenient approach for HLA-Cw DNA typing, allowing the definition of serologically undetectable alleles, and will contribute to the evaluation of the biological role of the HLA-C locus.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=41055Documentos Relacionados
- A novel method for simultaneous high resolution identification of HLA-A, HLA-B, and HLA-Cw alleles.
- Cryptosporidium parvum Mixed Genotypes Detected by PCR-Restriction Fragment Length Polymorphism Analysis
- Molecular typing of Borrelia burgdorferi sensu lato by PCR-restriction fragment length polymorphism analysis.
- Identification of Staphylococcus spp. by PCR-Restriction Fragment Length Polymorphism of gap Gene
- Molecular typing of Borrelia burgdorferi from Lyme disease patients by PCR-restriction fragment length polymorphism analysis.