联合应用PCR/SSCP和PCR/SSO提高HLA基因分型的精确性

PCR/SSCP used as a complementary method for HLA oligotyping

ＰＣＲ／ＳＳＯ分析７０例无血缘关系湖南汉族正常人ＨＬＡＤＰＡ１位点等位基因多态性，并对全部样本ＨＬＡＤＰＡ１基因第二外显子作银染ＰＣＲ／ＳＳＣＰ分析。发现ＰＣＲ／ＳＳＯ误将一例ＤＰＡ１＊０１／０２杂合子定为单一的ＤＰＡ１＊０１基因；经ＰＣＲ／ＳＳＣＰ完善获得湖南汉族正常人ＤＰＡ１＊０１，０２基因频率值分别为０．２６３２和０．７３２７；ＰＣＲ／ＳＳＣＰ法显示湖南汉族人ＤＰＡ１＊０２基因由单一亚型构成。结果表明，ＰＣＲ／ＳＳＣＰ可用于进一步分析ＰＣＲ／ＳＳＯ分型中杂交信号不典型或难以准确定型的样本，用于分析ＳＳＯ序列以外的ＨＬＡＤＮＡ多态性。

Seventy samples randomly selected from Hunan Han nationality normal individuals were typed for HLA DPA1 polymorphism by PCR/SSO with the primers and SSOs. Silver staining PCR/SSCP was also performed on the polymorphic second exon of all samples, which demonstrated the accurate gene frequencies of DPA1*01,02 to be 0.2632 and 0.7327, respectively. Using PCR/SSCP technique, we found that PCR/SSO mistyped a DPA1*01/02 heterozygote as single DPA1*01 gene because of weak hybridization signals. PCR/SSCP also revealed DPA1*02 in Hunan Han population to be composed of a single subtype. The results indicate that PCR/SSCP can be used to clarify the samples with atypical or irregular PCR/SSO hybridization patterns and to analyze the possible polymorphisms outside SSOs sequences. Thus this method may guarantee the accuracy of HLA oligotyping.