山西省非综合征型耳聋患者耳聋基因的检测分析

发布时间：2018-05-02 07:54

本文选题：耳聋 + 基因　；参考：《山西医科大学》2015年硕士论文

【摘要】：目的:本课题采用多种检测技术对非综合征性耳聋患者的致病基因进行筛查,统计常见基因的突变位点,寻找新的致病基因,为疾病的产前诊断、早期筛查及治疗提供依据;同时也为耳聋基因的诊断提供了更快速、简便、灵敏的方法。方法:由山西省太原市中心医院中心实验室收集山西省各地区聋哑学校的耳聋患者标本300例,年龄均在8-23岁之间。纳入标准为:除听力下降以外不伴有其他任何临床症状。通过对患者临床症状,检验结果及相关检查确诊并建立资料库。由患者本人或监护人签署知情同意书后采集外周血。采用基质辅助激光解吸电离飞行时间质谱法对4个常见基因二十个位点进行筛查,并统计突变位点,对未发生突变的标本选取10例用基因捕获+二代测序的方法进行检测,筛选出新的热点致病基因,并合成相应的引物,用Sanger测序的方法对剩余标本进行检测,寻找致病突变位点,用100例正常标本作为对照。结果:1.通过基质辅助激光解吸电离飞行时间质谱法对300例耳聋患者标本检测后,有152例发生突变。2.在剩余的148例标本中选取10例,经过基因捕获+二代测序的方法进行检测后,有3例发生TRIOBP基因的突变,2例发生TPRN基因的突变。3.通过对上述两个基因的相应外显子进行引物的设计与合成,对剩余的138例标本用Sanger测序进行检测,发现TRIOBP基因中,131例携带c.3559 TC(p.F1187L),11例携带c.3070 CA(p.P 1024 T)的突变,通过正常对照的验证,c.3559TCT可能是SNP位点,而c.3070 CA在正常对照组中未被发现,该位点能否导致耳聋,还有待进一步验证;TPRN基因未见突变。结论:1.GJB2、SLC26A4、GJB3、线粒体DNA这四个基因是山西省耳聋患者的主要致病基因,但各突变位点的携带率与全国其他地区有明显的差异;2.TRIOBP和TPRN基因的突变与非综合征性耳聋的发生有关;3.基质辅助激光解吸电离飞行时间质谱法和基因捕获+二代测序是两种诊断耳聋基因的快速、简便、灵敏的方法。
[Abstract]:Objective: to screen the pathogenetic genes of non-syndromic deafness patients by using various detection techniques, and to find out the mutation sites of common genes, so as to provide the basis for prenatal diagnosis, early screening and treatment of diseases. It also provides a more rapid, simple and sensitive method for the diagnosis of deafness gene. Methods: 300 deafness samples were collected from the central laboratory of Taiyuan Central Hospital of Shanxi Province, aged between 8 and 23 years. The inclusion criteria are: no clinical symptoms except hearing loss. The clinical symptoms, test results and related examinations were confirmed and the database was established. Peripheral blood was collected after informed consent was signed by the patient or guardian. Twenty loci of four common genes were screened by matrix assisted laser desorption ionization time of flight mass spectrometry, and mutation sites were counted. 10 samples without mutation were detected by gene capture and second generation sequencing. A new hot spot pathogenicity gene was screened out and the corresponding primers were synthesized. The remaining samples were detected by Sanger sequencing and the pathogenicity mutation sites were found. 100 normal samples were used as control. The result is 1: 1. 152 cases of deafness were detected by matrix assisted laser desorption ionization time of flight mass spectrometry. Among the remaining 148 samples, 10 cases were detected by gene capture second generation sequencing method, 3 cases of TRIOBP gene mutation occurred in 2 cases of TPRN gene mutation. 3. Through the design and synthesis of primers for the corresponding exons of the two genes mentioned above, the remaining 138 samples were detected by Sanger sequencing. The mutation of TRIOBP gene was found in 131cases with c.3559TCCnp.F1187L (11 cases carrying c.3070 CA(p.P 1024 T). It was confirmed by normal control that the SNP locus might be C3559TCT, but that c.3070CA was not found in the normal control group. Whether the locus could lead to deafness should be further verified that there was no mutation in the TPRN gene. Conclusion: 1. The four genes of GJB2, SLC26A4, GJB3 and mitochondrial DNA are the main pathogenetic genes in deafness patients in Shanxi Province, but there are significant differences between the carrying rate of each mutation locus and other regions in China. 2. The mutations of TRIOBP and TPRN genes are related to the occurrence of non-syndromic deafness. Matrix assisted laser desorption ionization time of flight mass spectrometry and gene capture sequencing are two rapid, simple and sensitive methods for the diagnosis of deafness genes.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R764.43;R440

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