【摘要】 目的研究云南籍葡萄糖-6-磷酸脱氢酶(G6PD)基因突变型特点,探讨检测G6PD缺乏症基因突变型的有效方法.方法应用硝基四氮蓝(NBT)纸片法进行G6PD缺乏症定性筛查,等位基因特异抗突变系统(ARMS),聚合酶链反应-单链构象多态性分析(PCR-SSCP),变性梯度凝胶电泳(DGGE)和DNA测序分析46例云南籍G6PD缺乏症患者的G6PD基因突变类型.结果46例样本经PCR-ARMS法发现nt-1388G→A 18例(39.13?),nt-1376G→T 2例(4.30?);经PCR-SSCP法发现有22例样本有电泳迁移率异常,其DNA测序结果与PCR-ARMS法的结果吻合,并发现2例nt-1311C→T;经PCR-DGGE法分析G6PD exon12发现有30例样本发现异常泳动条带,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G6PD exon2未发现有异常泳动条带的样本.结论(1)nt-1388G→A、nt-1376G→T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)在检测突变的PCR-ARMS法、PCR-SSCP法和PCR-DGGE法三种方法中,以PCR-DGGE法结合DNA测序,阳性检出率高,简便、快捷、灵敏、结果准确可靠.更多还原

【Abstract】 Objective To study the genotype of of gene mutations of Glucose-6-phosphate dehydrogenase(G6PD) in Yunnan province and to reseauh valid examination methods of gene mutations of G6PD.Methods NBT was used to qualitatively screen G6PD deficiency.46 G6PD deficiency patients were measured to detect genotypes of gene mutation by PCR-ARMS,PCR-SSCP,PCR-DGGE and DNA sequencing.Results In these 46 samples,18 samples were nt-1388G→A(39.13%) and 2 were nt-1376G→T by PCR-ARMS(4.3%).Abnormal electrophoretic mobility was found abnormal in 22 samples by PCR-SSCP and its result of DNA sequencing was the same with that by PCR-ARMS and 2 nt-1311C→T was found.Abnormal electrophoretic bands were discovered in 30 samples through analyzing exon 12 by PCR-DGGE,among which 26 samples were confirmed nt-1388G→T by DNA sequencing(56.52%),4 were nt-1376G→T(8.7%).It was the first time to detect nt-1388G→A in Lisu nationality of China.No abnormal band showed in 46 samples through analyzing G6PD exon 2 by PCR-DGGE.Conclusion(1)nt-1388G→A,nt-1376G→T are the principal genotypes of gene mutation of G6PD in Yunnan province.They are the most common genotypes of gene mutation in China and reveal that Chinese have the same ancestor.(2)Among PCR-ARMS,PCR-SSCP and PCR-DGGE,PCR-DGGE and DNA sequencing have higher positive detection rate and are simpler,quicker,more sensitive and reliable.更多还原