RESUMO
The role of major histocompatibility complex (MHC) class I chain-related gene A (MICA), a ligand of NKG2D, has been defined in human diseases by its allele associations with various autoimmune diseases, hematopoietic stem cell transplantation (HSCT) and cancer. This study describes a practical system to develop MICA genotyping by allele-specific primer extension (ASPE) on microarrays. From the results of 20 control primers, strict and reliable cut-off values of more than 30,000 mean fluorescence intensity (MFI) as positive and less than 3000 MFI as negative, were applied to select high-quality specific extension primers. Among 55 allele-specific primers, 44 primers could be initially selected as optimal primer. Through adjusting the length, six primers were improved. The other failed five primers were corrected by refractory modification. MICA genotypes by ASPE on microarrays showed the same results as those by nucleotide sequencing. On the basis of these results, ASPE on microarrays may provide high-throughput genotyping for MICA alleles for population studies, disease-gene associations and HSCT.
Assuntos
Alelos , Técnicas de Genotipagem , Antígenos de Histocompatibilidade Classe I/classificação , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Primers do DNA/química , Genótipo , Antígenos de Histocompatibilidade Classe I/genética , Antígenos de Histocompatibilidade Classe I/imunologia , Humanos , Reação em Cadeia da Polimerase/métodos , Polimorfismo GenéticoRESUMO
HLA-B*54:35 and -B*54:38 differ from HLA-B*54:01:01 in codons in exons 2 and 3.
Assuntos
Antígenos HLA-B/genética , Transplante de Células-Tronco Hematopoéticas , Doadores de Tecidos , Sequência de Bases , Códon/genética , Éxons/genética , HumanosRESUMO
HLA-B*15:400N differs from HLA-B*15:01:01:01 by nucleotide deletions from position 328 to 331 in exon 3.
RESUMO
HLA-B*58:01:20 differs from HLA-B*58:01:01:01 by a single synonymous nucleotide exchange at position 297 in exon 3.
RESUMO
HLA-A*26:132 differs from HLA-A*26:01:01:01 at nucleotides 269 and 346 in exons 2 and 3.
RESUMO
HLA-B*46:67 differs from HLA-B*46:01:01 in codons 94, 95, and 103 in exon 3.
Assuntos
Antígenos HLA-B/genética , Transplante de Células-Tronco Hematopoéticas , Teste de Histocompatibilidade/métodos , Doadores de Tecidos , Substituição de Aminoácidos , Sequência de Bases , Humanos , Homologia de Sequência , VoluntáriosRESUMO
HLA-A*33:110 differs from HLA-A*33:03:01:01 in a codon 149 in exon 3.