分子遗传学方法在遗传性角膜营养不良分类中的应用

Application of molecular genetic technique in classification of corneal dystrophy

目的探讨应用分子遗传学方法对角膜营养不良进行分类的途径。方法采用聚合酶链-单链构象多态性(PCR-SSCP)分析,对18例角膜营养不良的患者进行BIGH3基因的4、11、12号外显子的突变筛查,并对发现的异常泳动带进行DNA测序,以确定突变位点。结果18例角膜营养不良均有BIGH3基因突变,其中R555W突变型6例,A546D突变型6例,R124C突变型2例,T538P突变型2例,A546T突变型1例,P501T突变型1例。结论PCR-SSCP结合DNA测序的分子遗传学方法快速、简单且灵敏,以此为基础对遗传性角膜营养不良进行分类准确可靠。

Objective To study classification of hereditary corneal dystrophy by technique of molecualr genetics. Methods Polymerase chain reaction -single strand conformational polymorphism （PCR-SSCP） assay was performed to examine exons 4,11,12 of BIGH3 （Human transforming grwth factor beta induced） gene in 18 cases, and fragments with a mobility shift were sequenced to identify the gene mutations. Results Mutations in BIGH3 gene were found in all patients with corneal dystrophies. The mutation R555W in the BIGH3 gene was detected in six cases. The mutation A546D was detected in six cases. The mutation R124C was detected in two cases. The mutation T538P was detected in two cases. The mutation A546T was detected in one case. The mutation P501T was detected in one case. Conclusion The useful information is provided from PCR-SSCP and sequencing for making gene diagnosis and classification to hereditary corneal dystrophy.