HLA-A typing: comparison between serology, the amplification refractory mutation system with polymerase chain reaction and sequencing.
In this study we typed HLA-A polymorphisms by a new sequence-based typing (SBT) method, which involved one PCR reaction and four sequencing reactions covering exon 2 and exon 3. This method allowed complete identification of all known HLA-A alleles and revealed the presence of a new allele, named HLA-A*2608. We also introduced sequencing of exon 4 for some samples in order to discriminate the allelic pairs that are identical in exon 2 and 3, thus improving SBT resolution. Finally, we compared the results obtained by SBT with data obtained by serological typing and the amplification refractory mutation system (ARMS-PCR). Together, our results suggest that the SBT here described provides an optimal HLA-A typing technique that may be useful in selecting donor-recipient pairs in bone marrow transplantation between unrelated individuals.
Pera
C
C
National Institute for Cancer Research, IST, Advanced Biotechnology Center, Genova, Italy.
Delfino
L
L
Morabito
A
A
Longo
A
A
Johnston-Dow
L
L
White
C B
CB
Colonna
M
M
Ferrara
G B
GB
D000328:Adult; D000483:Alleles; D001483:Base Sequence; D001782:Blood Donors; D016026:Bone Marrow Transplantation / Q000276:immunology; D015343:DNA Probes, HLA; D005069:Evaluation Studies as Topic; D005091:Exons / Q000235:genetics; D005805:Genes, MHC Class I; D015234:HLA-A Antigens / Q000032:analysis* / Q000235:genetics; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D012698:Serologic Tests; D014019:Tissue Donors
D015343:DNA Probes, HLA; D015234:HLA-A Antigens; D011933:Reagent Kits, Diagnostic