PCR／SSCP技术在检测HLA基因点突变中的应用

聚合酶链反应－单链构象多态性（ＰＣＲ／ＳＳＣＰ）是根据核苷酸序列不同的单链ＤＮＡ，可形成不同构象，从而在不含变性剂的聚丙烯酰胺凝胶中泳动速度不同的原理，分离鉴定等长ＤＮＡ单链的方法，本文应用ＰＣＲ／ＳＳＣＰ方法检测了６０例无亲属关系个体白细胞抗原Ⅱ类基因（ＨＬＡ－ＤＱＡ１）５－上游调控区的等位基因多态性，并以ＰＣＲ直接测序证明每种ＳＳＣＰ带型都与不同位置单个碱基取代相关联。

Detection of HLAGene Mutation by PCR/SSCP Analysis

he technique for analyzing signle-stranded conformational polymorphisms(SSCP)is basedon the principle that the conformation of a single-stranded DNA molecule depends upon its nu-cleotide sequence.Even a point mutation can alter the conformation,causing a detectable shift inelectrophoretic mobility by natural polyacrylamide electrophoresis.In this study.we have analyzedthe polymorphism in HLA-DQA1 regulatory region by PCR/SSCP.Seven SSCP patterns havebeen found.on the basis of PCR-direct sequencing.it was demostrated that the difference ofSSCP patterns resulted from a single nucleotide substitution at a different position.