广西β地中海贫血基因突变类型SSCP分析

SSCP ANALYSES FOR THALASSEMIA GENE MUTATIONS IN GUANGXI

目的：建立一种简便易行，准确可靠，易于推广应用的β地中海贫血基因诊断方法。同时探讨单链ＤＮＡ碱基构成与最适ＳＳＣＰ电泳温度（Ｔｓ）的相关性。方法：应用巢式ＰＣＲ方法扩增β珠蛋白基因，然后用自行改良的ＳＳＣＰ方法对ＰＣＲ扩增片段进行分析。结果：１２种已知β地中海贫血突变均被检出。结论：改良的ＳＳＣＰ方法简便易行，准确可靠，适于推广应用。Ｔｓ与胞嘧啶碱基（Ｃ）成正相关，与腺嘌呤碱基（Ａ）成负相关，由此得出经验公式：Ｔｓ＝１１×Ｃ／Ａ。

Objective: To establish a simple,accurate and reliable gene diagnostic method and study the correlation between the base constitution of single strand DNA and the optimal electrophoretic temperature(Ts)for SSCP Method: globin gene was amplified by nested PCR,and then,the amplified fragments were analyzed by SSCP method improved by us Result: All of 12 kinds of known thalassemia mutations in Guangxi could be detected by the improved SSCP method Conclusion: The improved SSCP method is simple and easy to be used and is applied in thalassemia gene diagnosesTs is positively correlative to base C and negatively correlative to ATs=11C/A in our system