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目的 检测16个家系20例性连锁Alport综合征患者COL4A5基因突变。方法 采用PCR-变性凝胶梯度电泳(DGGE)-直接测序法检测患者COL4A5基因中30个外显子及其相邻内含子区域(外显子1～25、31、32、41、50、51),另选取100例正常人外周血DNA作为对照。结果 共发现4种突变,包括1种位于1号外显子上的无义突变(266C→T谷氨酰胺22终止密码),1种位于31号外显子上的错义突变(2757G→T甘氨酸852缬氨酸),以及2种分别位于1、25号内含子区域的剪接突变(283+1G→T、2150+1G→T)。结论 COL4A5基因为性连锁Alport综合征的致病基因,突变类型多样,尚未发现热点突变。类似于外显子突变,内含子突变同样具有致病意义,患者临床症状典型。查阅基因库,此4种突变均为首次报道。
Objective To detect COL4A5 gene mutations in 20 patients with Alport syndrome in 16 families. Methods 30 exons and their adjacent intron regions of COL4A5 gene were detected by PCR-denaturing gel gradient electrophoresis (DGGE) -dem direct sequencing (exons 1 ~ 25,31,32,41,50, 51), another 100 normal human peripheral blood DNA as a control. RESULTS: Four mutations were found, including one nonsense mutation located on exon 1 (266C → T-glutamine 22 stop codon) and one missense mutation located on exon 31 (2757G → T glycine 852 Valine) and two splice mutations (283 + 1G → T, 2150 + 1G → T) located in intron 1,25, respectively. Conclusion The COL4A5 gene is a causative gene of sex-linked Alport syndrome. The mutation types are diverse and no hot-spot mutations have been found. Similar to exon mutations, intron mutations also have pathogenic significance, typical clinical symptoms. Check the gene pool, these four mutations were first reported.