RESUMO
A new allele, now named HLA-C*02:138, was discovered during testing of a registry donor for possible stem cell transplantation.
Assuntos
Doadores de Sangue , Antígenos HLA-C/genética , Transplante de Células-Tronco Hematopoéticas , Teste de Histocompatibilidade/métodos , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA/métodos , Adulto , Alelos , Sequência de Bases , Humanos , Masculino , Sistema de Registros , Homologia de Sequência , TransplantadosRESUMO
Phased sequencing identified the HLA-C*07:607 allele in an African-American patient and sibling donor.
RESUMO
Screening methods for unknown DNA sequence variations are laborious, expensive, and relatively insensitive. To evaluate the sensitivity and specificity of denaturing high-pressure liquid chromatography (DHPLC) screening for unknown protein C gene (PROC) mutations, we studied 31 PROC-deficient patients. Eleven amplimers containing 4 kb of the PROC gene and spanning all exons, splice junctions, and the putative promoter and 3'-untranslated regions were amplified by PCR for each patient. Each amplimer (n = 341) was sequenced with a fluorescence-based method, and screened by DHPLC. Sequencing identified 10 unique mutations and three polymorphisms. Combining all mutations and polymorphisms, 227 amplimers were homozygous wildtype, and 63 and 51 were heterozygous and homozygous mutant, respectively. DHPLC screening correctly identified all amplimers (100% sensitivity and specificity). DHPLC is a rapid, automated, sensitive and specific screening method for unknown mutations within the PROC gene, and may be a useful screening method for unknown mutations within other genes.
