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目的探讨中国人群中前庭水管扩大(enlarged vestibular aqueduct,EVA)患者SLC26A4基因突变图谱,为相关基因的筛查及临床应用奠定基础。方法采集95例EVA核心家系,募集84名听力正常者及46名内耳结构正常的听力损失患者作为对照。提取基因组DNA,应用聚合酶链反应(PCR)的方法扩增SLC26A4基因的21个外显子,纯化PCR产物后直接测序,使用DNAStar及BioEdit序列比对软件分析SLC26A4基因的突变位点,利用Clustal软件进行同源氨基酸序列比对分析。结果在95例EVA核心家系中,93例发生了SLC26A4基因突变,约占97.9%(93/95)。其中双等位基因突变占88.4%(84/95),单等位基因和未发现突变的家系分别占9.5%和2.1%。93个家系中共发现38种突变,包括23种国际上尚未报道的突变,15种已报突变(5种仅报道于中国家系)。IVS7-2A>G是所有突变中最常见的突变,其在所有突变体中约占57.63%(102/177),75(75/95)个家系发生了此种突变。结论95例中国EVA核心家系的SLC26A4基因调查中,发现了一个中国特异性的SLC26A4基因突变谱:97.9%的中国EVA患者均可检测到SLC26A4基因的突变,双等位基因突变率约为88.4%;23种新的突变和5种仅报道于中国家系的突变在其他人群未曾检测到;IVS7-2A>G为中国人群前庭水管扩大患者最常见的突变。
Objective To investigate the mutation map of SLC26A4 gene in patients with enlarged vestibular aqueduct (EVA) in Chinese population and lay the foundation for the screening and clinical application of related genes. Methods Ninety-five core family members of EVA were recruited, 84 normal hearing patients and 46 normal hearing loss patients with inner ear were recruited as controls. Genomic DNA was extracted and 21 exons of SLC26A4 gene were amplified by polymerase chain reaction (PCR). The PCR products were purified and sequenced directly. The site of SLC26A4 gene mutation was analyzed by DNAStar and BioEdit software. Clustal Software homologous amino acid sequence alignment analysis. Results In 95 cases of the nuclear family of EVA, 93 cases of SLC26A4 gene mutation occurred, accounting for 97.9% (93/95). Binary alleles accounted for 88.4% (84/95), families with single allele and no mutation found 9.5% and 2.1% respectively. Thirty-eight mutations were found in 93 families, including 23 internationally unrecognized mutations and 15 reported mutations (5 reported only in Chinese families). IVS7-2A> G is the most common mutation among all mutations, accounting for about 57.63% (102/177) in all mutants and 75 (75/95) of such families occurring in this mutation. Conclusion A SLC26A4 gene mutation was found in 95 cases of Chinese EVA nuclear pedigree. A mutation of SLC26A4 gene was detected in 97.9% of Chinese patients with EVA, with a double-allele mutation rate of 88.4% ; 23 new mutations and 5 mutations reported only in Chinese families were not detected in other populations; IVS7-2A> G was the most common mutation in patients with vestibular aqueduct enlargement in China.