PCR-SSCP技术在Anderson Fabry病诊断中的应用

PCR-SSCP: A sensitive screening strategy for Anderson Fabry disease

目的:探讨聚合酶链反应-单链构像多态性分析对于筛选Anderson Fabry病患者的应用价值。方法:对52例肥厚型心肌病患者GLA基因的7个外显子行聚合酶链反应,其扩增产物行单链构像多态性分析。对有条带异常者的所有产物行测序分析,并测定其α-半乳糖苷酶A活性。对确诊的患者,以同样的方法筛选其家系成员。结果:1例患者的第一外显子的第1170位碱基由G突变为A。其α-半乳糖苷酶A活性明显低下,证实为该病患者。该突变使5′端非蛋白质编码区序列发生改变,通过影响mRNA的翻译过程而导致该病。结论:聚合酶链反应-单链构像多态性分析可作为Anderson Fabry病的有效筛选手段。

Objective： To analyze the value of polymerase chain reaction and single strand conformation polymorphism gel analysis in screening of Anderson Fabry disease. Methods： Specimens of peripheral blood were collected from 52 Chinese patients who had been diagnosed as hypertrophic eardiomyopathy. Single.strand conformation polymorphism gel analysis of the polymerase chain reaction amplified products was conducted to search for mutations in the seven exons of GLA gene. The activity of α-galactosidase A was measured, and the relatives of patients with Anderson Fabry disease were screened in the same way. Results： A novel mutation was identified in the first exon of one patient, a missense mutation （G→A） at nueleotide 1170. His low α-galactosidase A activity confirmed the diagnosis of Anderson Fabry disease. The mutation caused changes of 5＇ untranslated region in mRNA and affected the activity of α-galactosidase A through hindering the translation of mRNA. Conclusion： PCR-SSCP can be used as a sensitive screening strategy for Anderson Fabry disease.