本文選題：耳聾 + 基因�。� 參考：《山西醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的:本課題采用多種檢測技術(shù)對非綜合征性耳聾患者的致病基因進行篩查,統(tǒng)計常見基因的突變位點,尋找新的致病基因,為疾病的產(chǎn)前診斷、早期篩查及治療提供依據(jù);同時也為耳聾基因的診斷提供了更快速、簡便、靈敏的方法。方法:由山西省太原市中心醫(yī)院中心實驗室收集山西省各地區(qū)聾啞學(xué)校的耳聾患者標本300例,年齡均在8-23歲之間。納入標準為:除聽力下降以外不伴有其他任何臨床癥狀。通過對患者臨床癥狀,檢驗結(jié)果及相關(guān)檢查確診并建立資料庫。由患者本人或監(jiān)護人簽署知情同意書后采集外周血。采用基質(zhì)輔助激光解吸電離飛行時間質(zhì)譜法對4個常見基因二十個位點進行篩查,并統(tǒng)計突變位點,對未發(fā)生突變的標本選取10例用基因捕獲+二代測序的方法進行檢測,篩選出新的熱點致病基因,并合成相應(yīng)的引物,用Sanger測序的方法對剩余標本進行檢測,尋找致病突變位點,用100例正常標本作為對照。結(jié)果:1.通過基質(zhì)輔助激光解吸電離飛行時間質(zhì)譜法對300例耳聾患者標本檢測后,有152例發(fā)生突變。2.在剩余的148例標本中選取10例,經(jīng)過基因捕獲+二代測序的方法進行檢測后,有3例發(fā)生TRIOBP基因的突變,2例發(fā)生TPRN基因的突變。3.通過對上述兩個基因的相應(yīng)外顯子進行引物的設(shè)計與合成,對剩余的138例標本用Sanger測序進行檢測,發(fā)現(xiàn)TRIOBP基因中,131例攜帶c.3559 TC(p.F1187L),11例攜帶c.3070 CA(p.P 1024 T)的突變,通過正常對照的驗證,c.3559TCT可能是SNP位點,而c.3070 CA在正常對照組中未被發(fā)現(xiàn),該位點能否導(dǎo)致耳聾,還有待進一步驗證;TPRN基因未見突變。結(jié)論:1.GJB2、SLC26A4、GJB3、線粒體DNA這四個基因是山西省耳聾患者的主要致病基因,但各突變位點的攜帶率與全國其他地區(qū)有明顯的差異;2.TRIOBP和TPRN基因的突變與非綜合征性耳聾的發(fā)生有關(guān);3.基質(zhì)輔助激光解吸電離飛行時間質(zhì)譜法和基因捕獲+二代測序是兩種診斷耳聾基因的快速、簡便、靈敏的方法。
[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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R764.43;R440

【參考文獻】

相關(guān)期刊論文 前4條

1 劉芳;劉麗英;劉曉放;;有機質(zhì)譜新技術(shù)在現(xiàn)代科研中的應(yīng)用[J];黑龍江醫(yī)藥;2010年01期

2 李蘊;陳向平;陶錚;吳皓;;中度以上聽力障礙患兒助聽器或人工耳蝸植入后語言發(fā)育狀況分析[J];聽力學(xué)及言語疾病雜志;2010年02期

3 周永安;王湘;馬云霞;栗向韶;張景萍;曹淑琴;王月;薛梅輕;周麗英;張全斌;葉松華;;遺傳性非綜合征性常見耳聾基因診斷的研究[J];中國優(yōu)生與遺傳雜志;2011年07期

4 孫捷;韋桂玲;陳俞;張華;;86例非綜合征型耳聾患者耳聾基因芯片檢測結(jié)果分析[J];中華耳科學(xué)雜志;2013年02期

相關(guān)博士學(xué)位論文 前1條

1 陳華勇;人工合成寡核苷酸的液相色譜—質(zhì)譜研究[D];中國科學(xué)院研究生院（廣州地球化學(xué)研究所）;2005年

，

本文編號：1833030