RESUMO
The increasing demand for allele-level human leukocyte antigen (HLA) typing has led the sequence-based typing (SBT) to become the preferred method. In turn, the steady increase in the number of HLA alleles driven by the adoption of SBT as the ultimate typing method leads to the ever increasing number of cis/trans ambiguities. Over the last few years, additional sequencing with the commercially available group-specific sequencing primers (GSSPs) has replaced sequence-specific primer-polymerase chain reaction and group-specific amplification as the means of resolving cis/trans ambiguities in many laboratories. Here we summarize our 3-year experience in designing and utilizing GSSPs for resolution of HLA class I ambiguities. The panel of GSSPs used in our laboratory includes 14 primers for HLA-A, 18 for HLA-B, and 13 primers for HLA-C. The panel resolves 99.9% of all ambiguities.
Assuntos
Primers do DNA , Genes MHC Classe I/genética , Teste de Histocompatibilidade/métodos , Análise de Sequência de DNA/normas , Alelos , Sequência de Bases , Primers do DNA/genética , Sondas de DNA de HLA/análise , Sondas de DNA de HLA/genética , Reações Falso-Positivas , Humanos , Dados de Sequência Molecular , Estudos Retrospectivos , Análise de Sequência de DNA/métodos , Homologia de Sequência , Software , Especificidade por Substrato/genética , Especificidade por Substrato/imunologiaRESUMO
Twenty new human leukocyte antigen C alleles identified in 2200 minority individuals characterized by sequence-based typing.
