摘要
目的比较异源双链、单链构像多态（ＳＳＣＰ）、异源双链ＳＳＣＰ法筛查突变的效率，寻找简便有效的基因突变筛查方法。方法应用聚合酶链反应（ＰＣＲ）扩增２３个已知的杂合子突变，分别以异源双链、ＳＳＣＰ、异源双链ＳＳＣＰ法分析突变，对比各方法对突变的检出情况。结果２３个杂合子突变中，异源双链法、ＳＳＣＰ法与异源双链ＳＳＣＰ法检出突变分别为２２个（９６％）、２０个（８７％）和２３个（１００％）。异源双链法结果比ＳＳＣＰ法结果稳定，重复性好，两种方法之间有一定互补性，异源双链ＳＳＣＰ法兼有两种方法的特点。结论异源双链ＳＳＣＰ法突变检出率高，经济、简便，值得在基因突变筛查中推广应用。
Objective To compare the effectiveness of heteroduplex, SSCP and heteroduplexSSCP analyses in mutational screening. Methods DNA samples with 23 known heterozygous mutations were amplified by PCR. The PCR products were then analyzed by heteroduplex, SSCP and heteroduplexSSCP analyses separately to testify the effectiveness of each method.Results Of the 23 mutations, 22 were detected by heteroduplex analysis (96%), 20 by SSCP analysis (87%) and 23 by heteroduplexSSCP analysis (100%). Results from heteroduplex analysis are more stable and repeatable than those of SSCP and they may complement each other. HeteroduplexSSCP analysis has the advantages of heteroduplex and SSCP analyses. Conclusion HeteroduplexSSCP analysis is simple, economical and more effective than SSCP or heteroduplex analysis alone and it may be widely applied in mutational screening.
基金
广东省自然科学基金
卫生部回国人员启动基金
关键词
DNA
基因突变
异源双链
单链构像多态
DNA mutational analysis Singlestranded conformational polymorphism Nucleic acid heteroduplexes
