PCR／SSCP技术在检测HLA基因点突变中的应用  

Detection of HLAGene Mutation by PCR/SSCP Analysis

作　　者：朱席琳[1] 余跃[1] 杜志荣[1] 周文郁[1] 邱长春[1] 

机构地区：[1]中国医学科学院基础医学研究所

出　　处：《基础医学与临床》1996年第1期53-56,共4页Basic and Clinical Medicine

摘　　要：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－ＤＯＡ１）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。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.

关 键 词：白细胞抗原 突变 聚合酶链反应 单链构象多态性 

分 类 号：R392.13[医药卫生—免疫学]