基因芯片联合DNA测序法在大前庭水管综合征患者基因诊断中的应用  

Application of DNA microarray and sequencing in genetic diagnosis of enlarged vestibular aqueduct syndrome

作　　者：朱发梅[1,2] 胡鹏[1] 赖若沙[1] 谢鼎华[1] 

机构地区：[1]中南大学湘雅二医院耳鼻喉科,长沙410011 [2]南华大学附属南华医院耳鼻喉科,衡阳421002

出　　处：《中华耳科学杂志》2011年第2期168-173,共6页Chinese Journal of Otology

基　　金：国家自然科学基金(30000094和30070807);湖南省十一五期间人工耳蜗植入援助计划资助研究项目

摘　　要：目的应用基因芯片联合DNA测序法对大前庭水管综合征患者进行基因突变检测,分析中国人患者的基因型及探讨其基因诊断策略。方法采集30例大前庭水管综合征患者的外周血,提取基因组DNA。耳聋基因芯片筛查SLC26A4基因的2个突变ⅣS7-2A>G和H723R,发现纯合突变或复合杂合突变即停止筛查。如未发现突变或发现单纯杂合突变则采用DNA测序法检测SLC26A4基因其余外显子,直至发现另一个突变。如仍未发现突变则检测FOXI1基因。结果在30例大前庭水管综合征患者中,基因芯片法联合DNA测序法共检出28例有SLC26A4基因突变(93.33%)。共发现16种突变类型,其中ⅣS7-2A>G突变的发生率最高,其次为H723R。新发现4种突变类型(G368X、ⅣS8-1G>T、ⅣS13+9C>T和Q696X),未发现FOXI1基因突变。结论在中国人大前庭水管综合征患者中,SLC26A4基因的ⅣS7-2A>G突变的发生率最高,其次为H723R。针对这两个突变热点的基因芯片适用于对中国人群进行SLC26A4基因突变的筛查。发现的4例新突变类型对大前庭水管综合征的病因研究和基因诊断具有重要意义。Objective To study genotypes and effective genetic diagnostic approaches in patients with enlarged vestibular aqueduct syndrome （EVAS） using DNA microarray in combination with DNA sequencing. Methods Thirty patients with EVAS from unrelated Chinese families underwent genetic examination. Genomic DNA extracted from the patients was subjected to allele-specific PCR and universal array to screen for the Ⅳ S7-2A 〉 G or H723R mutant. The samples that carried only one or none of the two mutant alleles were subjected to PCR and sequenced to detect other mu- tations in the SLC26A4 and FOXH genes. Results A total of 16 SLC26A4 mutations were detected in 28 patients. Among them, 4 mutations were novel （G368X, ⅣS8-1G〉T, ⅣS13＋gc 〉 T and Q696X）. No FOXI1 mutation was found. Ⅳs7-2A〉G was the most common mutation in these patients, followed by H723R. Conclusions Four novel mutations have been detected in a group of Chinese patients with EVAS. This finding helps further our understanding of the cause for EVAS. The high prevalence of ⅣS7-2A〉G and H723R among Chinese EVAS patients makes genetic screening using DNA microarray possible and practical.

关 键 词：前庭水管 基因 基因芯片 突变 

分 类 号：R764.43[医药卫生—耳鼻咽喉科] R446.7[医药卫生—临床医学]