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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 explore the application of new clinical classification based on molecular genetics in the clinical diagnosis of corneal dystrophy through the detection of related gene mutations. Methods: Twenty cases of corneal dystrophy patients and 2 cases of normal peripheral venous blood from 10 outpatients and ward admitted to our hospital were collected. The leukocyte DNA was extracted by rapid method and specific primers were synthesized. The 4th and 12th exon of BIGH3 gene PCR amplification, PCR amplification products were purified and sequenced. RESULTS: All patients with corneal dystrophy had BIGH3 gene mutations, of which 13 were R12 4H heterozygotes and were diagnosed as Avellino corneal dystrophy; 3 were R5 5 5W heterozygotes and were diagnosed as granular corneal dystrophy; 4 were R12 4C heterozygous, diagnosed as lattice corneal dystrophy type Ⅰ. None of the normal controls had a BIGH3 gene mutation. Conclusion: This study confirms that the new clinical classification based on molecular genetics makes diagnosis more accurate, and at the same time makes it possible to predict the occurrence, development and prognosis of diseases more accurately and to provide a more fundamental, Effective treatment includes gene therapy has laid a good foundation.