应用聚合酶链单链构象多态技术检测葡萄糖6磷酸脱氢酶基因突变

为了探索一种快速、灵敏及准确的检测红细胞葡萄糖６－磷酸脱氢酶（Ｇ６ＰＤ）基因点突变方法，我们采用单链构象多态技术（ＳＳＣＰ），对２０例Ｇ６ＰＤ变异型基因外显子２进行分析。发现有４例患者双链ＤＮＡ中有一条链出现泳动变位，明显慢于正常人及其他患者，用ＰＣＲ直接侧序表明在ｃＤＮＡ第９５位核苷酸发生碱基置换（Ｔ→Ｃ）。结果说明ＳＳＣＰ技术检测点突变具有广泛的应用前景，比现行的点突变检测方法简便易行。作者还对ＰＣＲ－ＳＳＣＰ的优点和所检测突变的特点进行了探讨。

PCR-SSCP, DNA direct sequencing analysis in de-tecting mutation in exon 2 of G6PD gene.

ereditary glucose-6-phosphate dehydrogenase(G6PD) in red blood cell was one of the most commongenetic diseases in South China. The research data ofG6PD gene showed that at least 6 point mutationswere responsible for various G6PD vanants inChinese. For developing a rapid, sensitive, and cffec-tive method to detect point mutation, we applied single-strand confonnation (SSCP) analysis for detectionof mutation in exon 2 of G6PD gene of 20 cases ofG6PD variants. Four of them were found that mobili-ty shift band in one of two single strands DNA isslower than other individuals. PCR direct sequencingfor these 4 samples were given a base sustitution (T toC) at nucleotide 95 of cDNA. The results indicatedthat this technique is very simple, sensitive, and usefulover other methods of detecting point mutation. Theexperimental conditions of PCR-SSCP and featuresof this mutation were discussed at the same time.