葡萄糖-6-磷酸脱氢酶cDNA C1311T的多态性

Studies on a G6PD polymorphic site,cDNA C1311T

目的将突变特异性扩增系统（ＡＲＭＳ）法引入葡萄糖６磷酸脱氢酶（Ｇ６ＰＤ）Ｃ１３１１Ｔ突变型的检测，并利用它调查Ｃ１３１１Ｔ在正常人群中的发生频率，探讨此突变伴发的ＩＶＳ１１Ｃ９３Ｔ是否与酶活性降低有关。方法测序检测Ｇ６ＰＤ缺乏症患者中的Ｃ１３１１Ｔ突变，利用已证实为Ｃ１３１１Ｔ标本探索Ｃ１３１１Ｔ的ＡＲＭＳ检测法最佳条件，用此法来确定中国人中Ｃ１３１１Ｔ的发生频率。再利用测序探讨ＩＶＳ１１Ｃ９３Ｔ是否是某些病例酶活性降低的原因。结果在４０例Ｇ６ＰＤ缺乏症患者中检测到４例Ｃ１３１１Ｔ突变，摸索了Ｃ１３１１ＴＡＲＭＳ法检测的最佳条件。在１０３名正常男性中检测到１９名Ｃ１３１１Ｔ。对３名具有Ｃ１３１１Ｔ的Ｇ６ＰＤ缺乏症标本进行了Ｇ６ＰＤＩＶＳ１１测序，在９３位均为正常碱基Ｃ。同时检测到１份正常标本的ＩＶＳ１１的９３位为Ｔ。结论ＡＲＭＳ法是一种简单、省时、准确的筛查Ｇ６ＰＤ基因已知点突变的方法，Ｃ１３１１Ｔ在中国南方正常人群中的发生频率为１８４％，ＩＶＳ１１Ｃ９３Ｔ不是Ｃ１３１１Ｔ伴ＩＶＳ１１Ｃ９３Ｔ病例酶活性降低的原因，它极可能为Ｇ６ＰＤ基因的另一多态性位点。

Objective To use amplification refractory mutation system(ARMS) method to detect the G6PD cDNA C1311T mutation,estimate its frequency in a normal south Chinese population , and investigate whether IVS 11 C93T mutation is the cause of certain G6PD deficient cases.Methods DNA sequencing was used to confirm the C1311T and the IVS 11 C93T mutations. ARMS was set up to detect the C1311T and to estimate its frequency.Results Three cases of C1311T mutation were found in 40 G6PD deficient samples.The optimal condition for ARMS was established. Using this method, 19 cases of C1311T were detected in 103 normal men, and the frequency of this polymorphic mutation was estimated to be 18 4% in southern Chinese population. Four cases with G6PD deficiency were demonstrated to be C at the IVS 11 93 position. At the same time, a case of IVS 11 93 C→T was found in a normal man.Conclusions ARMS is a simple,time saving, and reliable method for detecting known G6PD gene point mutation.The frequency of C1311T in a normal south Chinese population is 18 4%.IVS 11 C93T might be another polymorphic site of the G6PD gene, and it is not the cause of enzyme deficiency in certain G6PD deficient cases.