Ambiguity results distribution and its solutions of HLA-A, B and DRB1 sequence-based typing / 重庆医学

Objective To investigate the ambiguity results distribution of HLA-A,B and DRB1 gene sequence-base typing in Guangxi population and to propose the way to resolve.Methods HLA-A,B and DRB1 genes of 1 000 donors in the Guangxi branch bank of China'bone marrow bank were genotyped by PCR-SBT,and then the ambiguity results distribution of the three loci was analyzed.The typing ambiguities resultswere resolved by high-resolution polymerase chain reaction-sequence-specific primers(PCR-SSP) and group specific sequencing primer(GSSP) methods,respectively.Results Among 1 000 samples,at least 1 locus in HLA-A,B and DRB1 genes in 96.7％ samples appeared the ambiguity results,in which the proportions of HLA-A,B and DRB1 loci appearing ambiguity results were 65.7 ％,58.8 ％ and 77.2 ％ respectively.For the samples of detected ambiguity results,single using the GSSP method could resolve the ambiguity typing results of 87.37％ HLA-A,93.54％ HLA-B and 60.49％ HLA-DRB1,using high-resolution PCR-SSP could resolve the ambiguity typing results of 12.63 ％ HLA-A,4.76 ％ HLA-B and 15.29 ％ HLA-DRB1,and the rest 1.70 ％ HLA-B and 24.22 ％ HLA-DRB1 ambiguity results were resolved by both GSSP and high-resolution PCR-SSPs method.Conclusion GSSP and high-resolution PCR-SSPs methods have high abilities to solve HLA ambiguity results both locate inside and outside the sequencing region,respectively.GSSP and high-resolution PCR-SSPs methods are supplement for each other,which can effectively resolve the problem of ambiguity results in high resolution HLA typing.

Identification of a novel HLA allele HLA-B*46:01:18 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To report on a novel human leukocyte antigen (HLA) allele.</p><p><b>METHODS</b>Polymerase chain reaction-sequence based typing was used for routine HLA typing. For one sample, the result of B locus typing showed mismatch of one base with B*46:01:01, B*15:25:01 at locus 384. The group specific sequencing primers, which target at B*46 and B*15, were used to confirm the difference between the novel allele and the highest homologous allele.</p><p><b>RESULTS</b>The sequencing results showed that the highest homologous allele to the novel allele was B*46:01:01. The two sequences only differed for position 384 within the exon 3 (384G>T), which resulted in a codon change (GGG>GGT), though the amino acid sequence of the novel allele at position 104 was still Glycine (G). Investigation of the family showed that the novel allele was inherited from the father.</p><p><b>CONCLUSION</b>The novel HLA-B allele, discovered in ethnic Zhuangs from Guangxi, has been designated as HLA-B *46:01:18 by the World Health Organization (WHO) HLA Nomenclature Committee.</p>

Preliminary analysis on antigen polymorphism of HLA-B15 and HLA-B40 groups among Han population in Guangxi area / 国际检验医学杂志

Objective To investigate the allele and specificity distribution situation of HLA-B15 group and HLA-B40 group antigens among Han population in Guangxi area and to explore their possible influence on transplantation donors selection in clinic.Methods The blood samples of 1 644 Han donors in Guangxi region were performed the HLA-B genotyping by PCR-SBT,the frequencies of each allele were calculated by the direct computing method.The antigen specificity of various alleles were analyzed,then the gene frequencies of HLA-B15 and HLA-B40 groups were compared with those from other populations.Results The gene frequency at HLA-B locus in 1 644 Han persons was inconsistent with the Hardy-Weinberg equilibrium(P<0.05).Fourteen alleles in HLA-B15 group were detected out,which belonged to 5 kinds of antigen specificity.In the HLA-B40 group,6 alleles were detected out,which belonged to two kinds of antigen specificity.Conclusion The antigen polymorphism of HLA-B15 and HLA-B40 groups among Han population in Guangxi area is close to that in southern Chinese Han populations,but which still keeps its characteristics of Guangxi area.