基因芯片在非综合征型耳聋基因突变中的应用

发布时间：2018-04-10 16:36

本文选题：耳聋 + GJB2基因　；参考：《青岛大学》2017年硕士论文

【摘要】：目的:探讨运用基因芯片和DNA测序技术检测非综合征型耳聋(non-syndromic hearing loss,NSHL)患者的基因突变类型的临床应用价值。方法:收集并分析患者一般情况、听力变动过程、加重或减轻的诱发因素、聋病史、家族史,检查全身情况以排除综合征型耳聋。采集非综合征型耳聋患者和健康体检者静脉血标本,提取DNA并进行PCR扩增,运用耳聋基因芯片技术确诊患者基因的突变类型,同时运用DNA测序方法进行结果验证。综合性分析我国各地区非综合征型耳聋患者的GJB2基因突变位点,研究对比不同实验中患者纳入标准、检测技术、实验数据之间差异性,制定我国统一的耳聋基因流行病学调查的规范化标准。选取健康体检者听力检测正常、没有遗传性听力下降家族史的体格正常的人群作为正常对照。结果:遗传性耳聋主要是指基因异常所致的耳聋,是人类最常见的感觉神经系统缺陷。这种疾病通过常染色体隐性,常染色体显性,X-连锁遗传和线粒体遗传方式遗传给下一代。在遗传性耳聋中,30%伴其他症状,称为综合征型耳聋;70%不伴其他症状,称为非综合征型耳聋(non-syndromic hearing loss,NSHL)。在GJB2(35del G、176dell6、235del C及299del AT)、GJB3(538CT)、SLC26A4(IVS7—2AG、2168AG)和线粒体DNA12Sr RNA(1555AG、C1494CT)这9个与非综合征型耳聋相关的基因位点中,检出GJB2基因235 del位点出现纯合突变,基因芯片的结果与测序结果完全一致,健康对照者中所有结果均为阴性;同时研究认为:NSHL患者人群中,GJB2基因编码区序列上共发现23种核苷酸改变,包括:5个框架移码突变、1个无义突变、15个错义突变及2个同义突变。GJB2基因突变在亚洲人遗传性耳聋中也有较高的携带比例,在中国人群的调查中,GJB2基因突变的主要方式为235de1C,是我国非综合征型耳聋人群中突最重要的致聋因素之一。结论:该非综合征型耳聋患者存在GJB2基因235 del位点纯合突变,且与扩增GJB2基因并进行DNA测序突变结果完全一致,基因芯片技术有助于诊断非综合征型耳聋。耳聋基因芯片法检测此类疾病,且联合分子流行病学调查以及通过产前基因诊断进行生育指导和干预均具有重要的临床意义。
[Abstract]:Objective: to investigate the clinical application value of gene mutation type in non-syndromic hearing (NHL) patients with non-syndromic deafness by gene chip and DNA sequencing.Methods: to collect and analyze the general situation of the patients, the process of hearing change, the aggravating or mitigating factors, the history of deafness, the family history, and to check the whole body condition to exclude the syndromic deafness.DNA was extracted from venous blood samples from non-syndromic deafness patients and healthy controls. The mutation type of genes was confirmed by using deafness gene chip technology. The results were verified by DNA sequencing method.The mutation sites of GJB2 gene in non-syndromic deafness patients in different regions of China were comprehensively analyzed, and the differences among the patient inclusion criteria, detection techniques and experimental data in different experiments were studied and compared.To establish a standardized standard for genetic epidemiological investigation of deafness in China.The healthy subjects with normal hearing test and no family history of hereditary hearing loss were selected as normal controls.Results: hereditary deafness mainly refers to deafness caused by genetic abnormality, which is the most common defect of human sensory nervous system.The disease is inherited to the next generation by autosomal recessive, autosomal dominant X-linkage and mitochondrial inheritance.30% of hereditary deafness were associated with other symptoms, 70% of which were called syndromic deafness without other symptoms, and were called non-syndromic hearing loss.All the results were negative in healthy controls and 23 nucleotide changes were found in the coding region of the GJB2 gene in the population with 1: NSHL.Among them, 5 frameshift mutations, 1 nonsense mutation, 15 missense mutations and 2 synonymous mutations. GJB2 gene mutations also have a high proportion of carriers in hereditary deafness in Asia.The main mutation pattern of GJB2 gene in Chinese population is 235de1C, which is one of the most important causes of deafness in non-syndromic deafness population in China.Conclusion: there is a homozygous mutation at the 235 del site of GJB2 gene in this non-syndromic deafness patient, which is consistent with the results of amplification of GJB2 gene and DNA sequencing. Gene chip technique is helpful for the diagnosis of non-syndromic deafness.It is of great clinical significance to detect the deafness disease by microarray method and to combine the molecular epidemiology investigation and prenatal gene diagnosis to guide and intervene the deafness disease.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R764.43

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