PCR—DGGE法研究葡萄糖-6-磷酸脱氢酶缺乏症基因突变

目的：应用PCR-DGGE法和DNA测序分析云南籍G6PD缺乏症患者基因突变类型和特点、方法应用硝基四氮唑蓝（NBT）纸片法进行G6PD缺乏症定性筛查，G6PD／6PGD比值法验证，应用PCR—DGGE法和DNA测序分析46例云南籍G6PD缺乏症患者基因突变类型和特点。结果：46例云南籍G6PD缺乏症样本中有30例经PCR—DGGE法分析G6PDexon12发现有异常电泳条带，DNA测序证实26例（56、52％）为nt-1388G→A，4例（8.7％）nt-1376G→T．而PCR—DGGE法分析G6PDexon2未发现有异常电泳条带的样本出现。结论：（1）nt-1388G→A（56．52％）、nt-1376G→T（8．7％）是云南省主要的基因突变型也是中国人中最常见的两种突变型，揭示中华民族有着共同的起源；（2）所检样本中未发现nt95A→G。（3）应用PCR—DGGE法结合DNA测序检测G6PD缺乏症患者的基因型，阳性检出率高，方法简便、快捷、灵敏、结果准确可靠。

Study on Gene Mutations of Glucose-6-phosphate Dehydrogenase Deficiency by PCR-DGGE

Objection： To study the genotype of Glucose-6-phosphate dehydrogenase deficiency of gene mutations in Yunnan province by polymerase chain reaction-denaturing gradient gel electrophorsis （PCR-DGGE） and DNA sequencing. Method： G6PD deficiency was detected by NBT qualitative. 46 cases of G6PD deficiency were measured to detect genotype of gene mutation by PCR-DGGE and DNA sequencing. Results： There was abnormal electrophoresis band discovered by analyzing exon 12 using PCR-DGGE in 30 cases. 26 cases （56.52%） were nt-1388G→T which were confirmed by DNA sequencing, and 4 cases （8.7%） were nt-1376G→T. No abnormal band of G6PD exon 2 was detected in 46 cases by PCR-DGGE. Conclusion： （1）nt-1388G→A （56.52%） and nt-1376G→T （8.7%） are the principal genotypes of gene mutation of G6PD in Chinese of Yunnan, and they are also the most common genotypes of gene mutation in Chinese, which revealed that Chinese have the same ancestor. （2） No nt95A→G were detected in 46 cases. （3） PCR-DGGE combined with DNA sequencing improve positive detection rate of genotype of G6PD deficiency, and it is a simple, sensitive, accurate and effective method for detecting gene mutations.