ABSTRACT
BACKGROUND: As all HLA class II genes, the DQ genes show their polymorphic variation mainly in the second exon, which encodes the first extracellular domain of the molecule. PCR-SSOP (Polymerase chain reaction-Sequence specific oligonucleotide probe) techniques were frequently used for HLA-DQA1 and DQB1 typing but certain alleles, DQA1*0101/0104/0105, *0302/0303, *0501/0505 and DQB1*0201/*0202, which differ from each other in segment other than exon 2, could not be unequivocally assigned. METHODS: To overcome this problem, we applied additional PCR-SSP (PCR-Sequence specific primer) method to analyze DQA1 exons 1, 3 and 4 and DQB1 exon 3. And we investigated the distributions and haplotypes of HLA-DRB1, DQA1 and DQB1 alleles in 406 unrelated Korean healthy individuals. RESULTS: Using this method the indistinguishable alleles of DQA1 and DQB1 in PCR-SSOP were typed definitively. We also found several important associations between DQA1 and DQB1 alleles in the Korean population; DQA1*0101-DQB1*0501, DQA1*0104-DQB1*0502 or -*0503, DQA1 *0105-DQB1*0501, DQA1*0302-DQB1*0303, DQA1*0303-DQB1*0401 or -*0402, DQA1 *0501-DQB1*0201, DQA1*0505-DQB1*0301, and DQA1*0201-DQB1*0202. The haplotypes of DRB1-DQA1-DQB1 associated with DQA1*01, *03, *05, and DQB1*02 subtypes were investigated. Several haplotypes associated with these alleles were observed in the Korean population. CONCLUSION: Our results can be helpful to find potential unrelated donors for bone marrow registries and study the HLA-associated disease and anthropology at high-resolution allelic level.
Subject(s)
Humans , Alleles , Anthropology , Bone Marrow , Exons , Genes, MHC Class II , Haplotypes , HLA-DRB1 Chains , Registries , Unrelated DonorsABSTRACT
PURPOSE: HLA (human leukocyte antigen)-class I genes are highly polymorphic, play many roles in organ and bone marrow transplantation. HLA-B is the most polymorphic class I locus with 414 alleles. HLA-class I typing, which is based on serologic method, has been used until recently. The development of molecular biological techniques make it possible to define the genotypes of HLA genes. METHODS: Analyses of HLA-B genotyping on 1,000 UCB (Umbilical Cord Blood) samples which were considered to be HLA-B homozygote or blank were performed by ARMS-PCR (Amplification Refractory Mutation System-PCR) method and direct sequencing. RESULTS: We could identify HLA-B*5001 which was known to be absent in Koreans. CONCLUSION: It is strongly suggested that HLA-B homozygote should be confirmed to the DNA level especially in cases of donor selection for the unrelated bone marrow transplantation.
Subject(s)
Alleles , Bone Marrow Transplantation , DNA , Donor Selection , Fetal Blood , Genotype , HLA-B Antigens , Homozygote , LeukocytesABSTRACT
The thirteen DRB1, 6 DQA1, and 5 DQB1 alleles were defined in 362 healthy Korean controls using reverse dot blot hybridization method. The twenty-four immobilized SSOs for DRB1, 8 for DQA1, and 6 for DQB1 were used for this study. The frequencies of genotypes were DRB104 (17.1'Yo), '09 (13.1%), and '13 (11.6%); DQA1'01 (46.7%), 03 (30.8%), and '05 (11.7%); DQB1*03 (39.5%), '06 '(29.8%), and 05 (16.0%). ...continue...