RESUMEN
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.
Asunto(s)
Cartilla de ADN , Genes MHC Clase I/genética , Prueba de Histocompatibilidad/métodos , Análisis de Secuencia de ADN/normas , Alelos , Secuencia de Bases , Cartilla de ADN/genética , Sondas de ADN de HLA/análisis , Sondas de ADN de HLA/genética , Reacciones Falso Positivas , Humanos , Datos de Secuencia Molecular , Estudios Retrospectivos , Análisis de Secuencia de ADN/métodos , Homología de Secuencia , Programas Informáticos , Especificidad por Sustrato/genética , Especificidad por Sustrato/inmunologíaRESUMEN
Twenty new human leukocyte antigen C alleles identified in 2200 minority individuals characterized by sequence-based typing.