河北省沧州地区非综合征性耳聋患者易感基因筛查分析

发布时间：2018-07-16 22:06

【摘要】：耳聋可导致与他人交流、沟通障碍,是最常见先天性疾患之一,本世纪初在全国范围内进行第二次大规模的残疾人取样调查分析结果表明目前听障患者数量可达2780万,当中约27%(127万)存在语言残疾,居总残障人士首位。国内出生缺陷性疾病之中先天性听力障碍最为常见。根据统计每一年有3万余儿童成为残障人士队伍中的新成员,这些儿童年龄均不超过7岁。还有迟发型或(和)药物诱发性耳聋患者人数逐年增高。人类基因组计划的完成使得基因相关领域的科学技术的发展,随之而来的有关先天性遗传相关性聋病患者的基因学研究同样获得重大突破,目前普遍认为耳聋患者的病因越来越偏向于遗传因素,此部分患者占耳聋患者人数的66%。80%的听力损失患者除耳聋外不伴随其他症状,称之为非综合症性耳聋(nonsyndromic hearing impairment.NSHI)。多个基因与遗传性听力障碍存在相关,其中大部分遗传性耳聋的相关基因定位于GJB2、SLC26A4和线粒体DNA病理突变。近年通过国内大规模耳聋基因筛查工作的开展发现在国民听力正常人群中存在较高的聋病相关易感基因突变的携带率,这部分患者将这种基因遗传给下一代,大大增加他们生育聋儿的风险。因此早诊断,早干预,对减少本地区先天性耳聋患者的出生率,及预后、建议选择配偶等方面具有非常重要的意义。目的:调查河北省沧州地区非综合征性耳聋患者聋病易感基因突变分子机理。方法:河北省沧州特殊教育学校学生及沧州市听力诊疗中心就诊的散发病例中诊断为先天性非综合征性耳聋的患者共358例,告诉患者及其家长本次调查研究目的,并填写问卷调查表及签署知情同意书;检查每位患者行全身体格检查除外有其他症状的综合征性耳聋;给每位患者进行纯音电听阈值检查(GSI-61纯音测听仪)及声导抗(Madsen声导抗检测仪),3岁以下耳聋患者行脑干诱发电位。抽出每位患者外周静脉血5ml,给予乙二胺四乙酸二钾抗凝,放入4℃冰箱中进行保存。全血样品完全解冻后,按照指示剂说明提取DNA。用紫外分光光度计测定样品的浓度和纯度。参照试剂盒说明书进行PCR扩增,杂交和洗涤后检测GJB2、SLC26A4、线粒体DNA 12SrRNA、GJB3聋病相关的4个基因的9个突变位点:GJB2c.235delG、GJB2 c.235delC、GJB2 c.176del16、GJB2 c.299del AT、SLC26A4c.2168AG、SLC26A4 IVS7-2AG、线粒体DNAC1494T和线粒体DNA A1555G,GJB3 c.538CT。结果:358名受试者中有占总人数19.5%(70人)检测出耳聋相关基因突变,其中GJB2基因突变患者为39人,占总人数的10.89%,GJB2235delc纯合突变患者为18人,299delAT纯合突变患者为3人,del235c位点突变单杂合突变患者为9人,复合杂合突变患者为9人;299delAT检出率占25%,176del16检出率占5.12%,235delC检出率为74.33%;SLC26A4突变患者28人,占7.82%,IVS7-2AG纯合突变患者为4人,IVS7-2AG单杂合突变患者为20人,复合杂合突变患者为4人。线粒体DNAA1555G均质突变患者仅为1人,占总人数0.27%。GJB3突变患者为2人占总人数0.54%。将受试者根据就诊方式不同分为特教组及散发组,特教组致病基因总检出率为15.35%(37/241),散发组总检出率为28.20%(33/117),P0.01,两组间存在差异,GJB2,SLC26A4,线粒体DNA,GJB3基因突变在特教组检出率分别为(10.3%,4.9%,0,0)。而散发组上述基因检出率为(11.7%,13.6%,0.85%,1.75%),GJB2基因突变率在两组间不存在差异P0.05,而SLC26A4基因突变率在两者间存在差异,P0.01,具有统计学意义。结论:通过检测聋病基因检测位点,进行聋病分子诊断,对本地区预防耳聋发生、婚育、指导康复及评估预后提供依据。聋病相关基因突变检出率普遍低于全国水平,或许存在环境或其他遗传因素在本区域内发挥作用。
[Abstract]:Deafness can lead to communication with others, communication barriers, and one of the most common congenital disorders. At the beginning of this century, second large-scale persons with disabilities sampling survey showed that the number of hearing impaired patients reached 27 million 800 thousand, of which about 27% (1 million 270 thousand) were handicapped and the first. Congenital hearing impairment is the most common in the disease. According to statistics, more than 3 million children have become a new member of the disabled people each year. These children are not more than 7 years old. And the number of delayed or (and) drug induced deafness is increasing year by year. The completion of the human genome project makes the science and technology in the field of gene related. Development, the genetic study of patients with congenital genetic related deafness is also a major breakthrough. It is widely believed that the cause of the deafness is becoming more and more hereditary, and this part of the 66%.80% hearing loss patients, which is deafness, is not accompanied by other symptoms, and is called non syndrome. Nonsyndromic hearing impairment.NSHI. Multiple genes are associated with hereditary hearing impairment, and most of the genetic deafness related genes are located in GJB2, SLC26A4 and mitochondrial DNA pathological mutations. In recent years, the development of large-scale hearing loss screening in China has been found to be higher in people with normal hearing. The incidence of mutations in the susceptible genes of the deafness, which is inherited to the next generation, greatly increases their risk of having deaf children. Therefore, early diagnosis and early intervention are of great significance to reduce the birth rate and prognosis of the local deafness, and recommend the choice of spouses. Objective: To investigate Hebei Methods: 358 cases of congenital non syndromic deafness were diagnosed in the Cangzhou special education school and the hearing diagnosis center in Cangzhou, Hebei Province, and 358 cases with congenital non syndromic deafness were diagnosed in the Cangzhou special education school in Hebei province. The volume of the questionnaire and the signing of the informed consent; the general physical examination of each patient except for the syndrom deafness with other symptoms; the pure tone threshold examination (GSI-61 pure tone audiometry) and the acoustic conductance (Madsen acoustic conductivity detector) for each patient and the brainstem evoked potential of the deafness under 3 years of age. Vein blood 5ml, give ethylene diamine tetra acetic acid two potassium anticoagulant, put it in 4 centigrade refrigerator and store it. After complete thawing of the whole blood sample, the concentration and purity of the samples are determined by the ultraviolet spectrophotometer according to the indicator. PCR amplification, GJB2, SLC26A4, mitochondrial DNA 12SrRNA, GJB3 deafness after the hybridization and cleaning of the reagent box are amplified. 9 mutations of the 4 genes related to the disease: GJB2c.235delG, GJB2 c.235delC, GJB2 c.176del16, GJB2 c.299del AT, SLC26A4c.2168AG, SLC26A4 IVS7-2AG, mitochondrial DNAC1494T and mitochondrial DNA, which accounted for 19.5% (70) of the total number of mutations in the deafness related genes. The patients were 39, accounting for 10.89% of the total number, 18 GJB2235delc homozygous mutations, 3 homozygous mutations, 9 heterozygous mutations in del235c site and 9 in complex heterozygous mutations, 25% in 299delAT, 5.12% in 176del16, 74.33% in 235delC, 28 in SLC26A4, 7.82% in SLC26A4. The IVS7-2AG homozygous mutation was 4, the IVS7-2AG single heterozygous mutation was 20, the complex heterozygous mutation was 4. The mitochondrial DNAA1555G homozygous mutation was only 1, and the total number of 0.27%.GJB3 mutations was 2 of the total number 0.54%.. The subjects were divided into special and sporadic groups according to the different ways of visiting. The detection rate was 15.35% (37/241), the total detection rate of the sporadic group was 28.20% (33/117), and there were differences among the two groups, GJB2, SLC26A4, and mitochondrial DNA, and the detection rates of GJB3 gene mutations in the special group were (10.3%, 4.9%, 0,0), and the detection rates of the genes were (11.7%, 13.6%, 0.85%, 1.75%), and the mutation rate of GJB2 gene was not different P0.05 in the two groups. The mutation rate of SLC26A4 gene is different between the two and P0.01, which has statistical significance. Conclusion: the detection of deafness gene detection site and molecular diagnosis of deafness can provide a basis for prevention of deafness, marriage and breeding, guidance for rehabilitation and evaluation of prognosis. Environmental or other genetic factors play a role in the region.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R764.43

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