摘要
目的 :通过对相关基因突变的检测 ,探讨以分子遗传学为基础的新的临床分类方法在角膜营养不良临床诊断中的应用。方法 :取我院门诊及病房收治的角膜营养不良患者 2 0例及 10例正常人外周静脉血 2ml,采取快速法提取白细胞DNA ,合成特异引物 ,分别行BIGH3基因第 4及第 12外显子PCR扩增 ,将PCR扩增产物进行纯化和测序。结果 :所有角膜营养不良患者均有BIGH3基因突变 ,其中 ,13例为R12 4H杂合子 ,确诊为Avellino角膜营养不良 ;3例为R5 5 5W杂合子 ,确诊为颗粒状角膜营养不良 ;4例为R12 4C杂合子 ,确诊为格子状角膜营养不良Ⅰ型。所有正常对照者均无BIGH3基因突变。结论 :通过本研究证实 ,以分子遗传学为基础的新的临床分类方法使诊断更为准确 ,同时使更为准确地预测疾病的发生、发展及预后成为可能 ,并为今后进行更为根本、有效的治疗包括基因治疗打下了良好的基础。
Objective:To study the application of molecular genetics on the clinical diagnosis and classification of corneal dystrophies.Methods:2 ml peripheral venous blood was collected from 20 patients with corneal dystrophies and 10 normal subjects.Leucocytes DNA was extracted with standard method.With two pairs of oligonucleotide primers,exon 4 and exon 12 of BIGH 3 gene were amplified using the polymerase chain reaction.Amplified DNA fragments were purified and sequenced directly.Results:Mutations in BIGH 3 gene were detected in all the patients with corneal dystrophies.13 patients with Avellino corneal dystrophy had the missense mutation R124H in the BIGH 3 gene.3 patients with granular corneal dystrophy had the missense mutation R555W in the BIGH 3 gene.4 patients with lattice corneal dystrophy had the missense mutation R124C in the BIGH 3 gene.Conclusion:With molecular genetic analysis,proper diagnosis and classification of corneal dystrophies can be obtained.On the basis of such research,it will come true to predict occurrence,progress and prognosis of corneal dystrophy recently.Gene therapy will be performed in the near future.
出处
《眼科》
CAS
2003年第6期327-329,共3页
Ophthalmology in China
