【摘要】 目的分析SSP方法难以确定B* 48与B* 1568低分辨分型的原因。方法应用基因克隆和DNA测序技术对受检标本HLA-B基因第1-3外显子(Exon1-3)作核苷酸序列分析,通过序列比对寻找SSP分型困难的原因。结果该标本HLA-B基因Exon1序列与B*4801Exon1序列完全相同,与已公布的B* 15Exon1序列相比,第5位碱基由G→T,第11位碱基由C→T,第44、45位碱基由GA→CG。Exon3序列则与B* 1568完全相同,按WHOHLA命名该标本为B* 1568。结论部分PCR-SSP引物序列涉及Exon1区域,但有些等位基因在此区域的序列并未公布,因此可能出现分型困难。分析SSP分型中引物发生的疑难问题,有助于提高HLA分型水平。更多还原

【Abstract】 Objective To study the reason why PCR-SSP is difficult in determining some alleles at HLA-B locus.Methods DNA sub-cloning and sequencing were used to analyze the nucleotide sequence of HLA-B Exons 1、 2、 3 of samples. Sequence blasting was applied to work out the reason of unusual reaction pattern by SSP genotyping.Results The sample had the same sequence of Exons 2 and 3 as B*1568; the same sequence of Exon 1 as B*4801. However, Exon 1 sequence of the sample is different from the published Exon 1 of B*15 group at nt 5 G>T, nt 11 C>T, nt 44 and 45 GA>CG. And that was why SSP failed to determine the allele.Conclusion The difficulty in HLA genotyping by SSP resulted from the primers, which involved unknown sequence of Exon 1 at locus B in the studied sample.更多还原