本文選題：耳聾 + 基因芯片��； 參考：《揚(yáng)州大學(xué)》2012年碩士論文

【摘要】：耳聾,即指聽力障礙或聽力損失。是人們對聲音大小和辨識(shí)度下降的表現(xiàn)。耳聾是導(dǎo)致交流障礙最常見的疾病,對日常生活的影響較重,常常因聾致啞,對患者無疑又是雪上加霜。研究發(fā)現(xiàn)耳聾的病因主要由遺傳因素(60%)引起。而遺傳性聾分為非綜合征性耳聾(nonsyndromic hearing impairment,NSHI)和綜合征性耳聾(syndromic hearing impairment,SHI)。綜合癥性耳聾臨床表現(xiàn)形式多種多樣,非綜合癥性耳聾以單一神經(jīng)損害為主。非綜合征性耳聾(NSHI)以聽力損失為單一癥狀。NSHI具有高度遺傳異質(zhì)性,醫(yī)學(xué)遺傳專家估測可能有100多個(gè)基因位點(diǎn)與NSHI有關(guān)。按遺傳方式可分為：常染色體隱性遺傳約占70%,常染色體顯性遺傳約占22%,X連鎖約占1%,線粒體突變母系遺傳約占1%�；谌祟惢蚪M計(jì)劃的完成,耳聾基因在遺傳性耳聾中的研究也取得了突破性進(jìn)展。我國聾病分子流行病學(xué)調(diào)查顯示中國聾人群體中遺傳性耳聾比例高,且GJB2、GJB3, SLC26A4、線粒體基因突變是造成中國人耳聾的主要遺傳致病因素。運(yùn)用基因芯片篩查技術(shù)對GJB2, GJB3.SLC26A4.mtDNA基因進(jìn)行檢測,不但可以診斷70%的遺傳性耳聾患者的致聾原因,更為有再生育要求的耳聾夫婦生育聽力正常兒提供理論依據(jù)和科學(xué)保障。耳聾的遺傳咨詢因此具有重大的意義,可以科學(xué)合理的指導(dǎo)耳聾夫婦的婚育,減少耳聾患兒的出生率,減輕國家和耳聾家庭的經(jīng)濟(jì)負(fù)擔(dān)�；谏鲜�,本研究首先對揚(yáng)州地區(qū)中重度以上聽力障礙兒童進(jìn)行相關(guān)耳聾基因篩查,其次將基因篩查結(jié)果運(yùn)用于臨床,對有再生育要求的耳聾家庭進(jìn)行遺傳咨詢,為其生育聽力正常兒提供科學(xué)保障。 摘要 第一部分揚(yáng)州地區(qū)中度以上聽力障礙兒童的耳聾基因篩查研究 實(shí)驗(yàn)?zāi)康模簯?yīng)用耳聾基因芯片檢測技術(shù)對揚(yáng)州地區(qū)中度以上聽力障礙兒童進(jìn)行篩查,探討遺傳性耳聾基因在揚(yáng)州地區(qū)的分布規(guī)律。 實(shí)驗(yàn)方法：以自愿為原則,經(jīng)調(diào)查問卷研究及簽署知情同意書后,對來自揚(yáng)州市特殊教育學(xué)校的65名非綜合癥性耳聾學(xué)生(其中45名為先證者,設(shè)為實(shí)驗(yàn)組；其余20名無明顯遺傳性,設(shè)為對照組),進(jìn)行全身體格檢查及耳鼻喉�？茩z查,聽力學(xué)測試等證實(shí)為非綜合癥型耳聾(聽力下降為唯一臨床癥狀),提取外周靜脈血3ml,應(yīng)用小劑量全血基因組DNA提取試劑盒提取DNA,后經(jīng)基因聚合酶鏈(PCR)反應(yīng)擴(kuò)增檢測基因,并雜交,洗片等,后應(yīng)用晶芯“九項(xiàng)遺傳性耳聾基因”檢測試劑盒檢測受試者4個(gè)耳聾相關(guān)基因上的9個(gè)熱點(diǎn),包括GJB2(35delG,176de116bp,235delC,299_300del AT). GJB3(538CT,547GA)、SLC26A4(IVS7-2AG,2168AG)和線粒體DNA12s rRNA (A1555G).使用晶芯Lux Scan TM10KB微陣列芯片掃描儀以90的激光掃描強(qiáng)度和532nm激光波長行芯片掃描以及相應(yīng)的遺傳性耳聾基因芯片檢測判別系統(tǒng)進(jìn)行信號(hào)讀取和判斷。 實(shí)驗(yàn)結(jié)果：檢測人群中總陽性檢出率(至少攜帶一個(gè)突變等位基因)研究組61.54%(40/65),其中：GJB2235delC純和突變14例,235de1C雜和突變3例,235delC/299delAT復(fù)合雜合突變2例,176de116純和突變1例,299delAT純合突變2例,299delAT雜合突變1例,176del16/235delC雜合突變1例,GJB2突變陽性率占總突變的60.00%(24/40);SLC26A4ⅣS7-2AG純和突變2例,SLC26A4ⅣS7-2AG雜和突變7例,SLC26A4ⅣS7-2AG/SLC26A42168AG復(fù)合雜合4例,SLC26A42168AG雜和2例,SLC26A4突變陽性率占總突變的37.50%(15/40)；線粒體DNA12SrRNA A1555G均質(zhì)1例,突變陽性率占總突變的2.50%(1/40)；GJB3突變陽性率為0；5例未檢出上述突變基因；對照組20例；均未檢出突變基因。 實(shí)驗(yàn)結(jié)論：揚(yáng)州地區(qū)遺傳性耳聾具有較高的發(fā)病率,GJB2是主要的致聾基因,SLC26A4檢出率高于全國平均水平。 第二部分基于基因診斷的聾病遺傳咨詢 實(shí)驗(yàn)?zāi)康模和ㄟ^基因檢測結(jié)果并結(jié)合有再生育要求的耳聾夫婦的基因篩查結(jié)果,預(yù)測其再生育耳聾患兒的風(fēng)險(xiǎn)。 實(shí)驗(yàn)方法：在知情同意的情況下,遵循保密等原則,通過調(diào)查問卷及首胎基因檢測結(jié)果,應(yīng)用耳聾基因芯片技術(shù)對有再生育要求的夫婦進(jìn)行檢測并進(jìn)行遺傳咨詢。 實(shí)驗(yàn)結(jié)果：結(jié)合首胎患兒及其父母的基因篩查結(jié)果,該6對夫婦中；2對夫婦懷孕期間需行產(chǎn)前診斷：一對夫婦懷孕期間不需行產(chǎn)前診斷：3對夫婦懷孕期間不必行產(chǎn)前診斷。 實(shí)驗(yàn)結(jié)論：耳聾家族再生育聾兒概率高,其再生育前行耳聾基因檢測可以預(yù)知胎兒患病風(fēng)險(xiǎn)。
[Abstract]:Refers to the deaf, hearing impairment or hearing loss. Is the sound drop size and recognition performance. The most common cause of deafness is a communication disorder effect on daily life is heavy, often because of dumb, the patient is undoubtedly one disaster after another. The study found that the disease mainly due to deafness by genetic factors (60%). Genetic deafness is divided into nonsyndromic deafness (nonsyndromic hearing, impairment, NSHI) and syndromic deafness (syndromic hearing, impairment, SHI). The clinical manifestations of deafness syndrome varied, non syndrome deafness in a single nerve damage. Non syndromic deafness (NSHI) hearing for the loss of single symptom.NSHI has a high degree of genetic heterogeneity, genetic medicine experts estimate that there may be more than 100 loci associated with NSHI. According to the mode of inheritance can be divided into: autosomal recessive autosomal accounted for about 70% Dominant accounted for 22%, accounting for about 1% of the X chain, mitochondrial mutations in maternally inherited accounts for about 1%. of the completion of the human genome project based on the study of deafness gene in genetic deafness also made a breakthrough. The investigation of deafness molecular epidemiology in China showed that China deaf genetic deafness in a high proportion, and GJB2. GJB3, SLC26A4, mitochondrial gene mutation is the main cause of genetic deafness Chinese pathogenic factors. Using gene chip screening technology of GJB2, GJB3.SLC26A4.mtDNA gene detection, not only can diagnose 70% of genetic deafness patients for deaf, more have fertility requirements of deaf couples with normal hearing children and the scientific basis for protection genetic counseling for deafness. It is therefore of great significance to the guidance of scientific and rational deaf couples of marriage, reduce the birth rate of deaf children, and reduce the state's deafness family Family economic burden. Based on the above, this study first in the Yangzhou area of moderate to severe hearing impaired children by screening the deafness gene, then the gene screening results in clinical use, to have fertility requirements of deaf families for genetic counseling, provide a scientific guarantee for the normal hearing children. Family
abstract
The first part of the hearing loss gene screening of children with moderate hearing impairment in the first part of Yangzhou
Objective: to detect the distribution of hereditary deafness gene in Yangzhou area by screening deafness gene chip technology for children with moderate hearing impairment in Yangzhou area.
Methods: the experiment voluntarily, through questionnaire research and signed informed consent, 65 non syndrome deafness students from Yangzhou City special education schools (including the proband, 45 for the experimental group; the remaining 20 were hereditary, as control group), body physique check and ENT specialist examination, audiological testing confirmed the non syndromic deafness (hearing loss as the only clinical symptoms), peripheral venous blood 3ml extraction, application of small dose of whole blood genomic DNA extraction kit to extract DNA by polymerase chain reaction (PCR) amplification and hybridization, gene detection, processing etc. after the application of geedom "detection of nine genetic deafness gene detection kit subjects 9 hot 4 deaf related genes, including GJB2 (35delG, 176de116bp, 235delC, 299_300del, AT). GJB3 (538CT, 547GA), SLC26A4 (IVS7-2AG, 2168AG) and Mitochondrial DNA12s rRNA (A1555G). The use of Lux Scan TM10KB chip microarray scanner for signal read and judge with 90 laser scanning intensity and laser wavelength scanning for 532nm chip and the corresponding genetic deafness gene chip detection system.
Results: the total positive rate of test group (carry at least one mutant allele) study group 61.54% (40/65), including: GJB2235delC homozygous mutation in 14 cases, 3 cases of 235de1C mutation, 235delC/299delAT compound heterozygous mutation in 2 cases, and 1 cases of pure 176de116 mutation, 299delAT homozygous mutation in 2 cases 299delAT, a heterozygous mutation in 1 cases, 176del16/235delC 1 cases with heterozygous mutation and GJB2 mutation positive rate of the total 60% mutations (24/40); SLC26A4 IV S7-2AG homozygous mutation in 2 cases, SLC26A4 IV S7-2AG mutation in 7 cases, 4 cases of SLC26A4 IV S7-2AG/SLC26A42168AG compound heterozygous, SLC26A42168AG heterozygous SLC26A4 mutation positive and 2 cases. The total rate of mutation in 37.50% (15/40); 1 cases of mitochondrial DNA12SrRNA A1555G mutation positive rate of homogeneous, total 2.50% mutations (1/40); GJB3 mutation positive rate was 0; 5 cases were not detected in the mutant gene; 20 patients in control group were not detected; gene mutation.
Experimental conclusion: the incidence of hereditary deafness in Yangzhou is high, and GJB2 is the main deafness gene, and the detection rate of SLC26A4 is higher than the national average.
The second part of genetic counseling based on genetic diagnosis of deafness
Objective: to predict the risk of deafness in children with deafness through gene screening and genetic screening of deaf couples with reproductive requirements.
The experimental method: following the informed consent, we should follow the principles of confidentiality and so on. Through the questionnaire and first birth gene detection results, we used deafness gene chip technology to detect couples with reproductive requirements and conduct genetic counseling.
Experimental results: combined with the first child and their parents' genetic screening results, the 6 couples, 2 couples need prenatal diagnosis during pregnancy: one couple does not need prenatal diagnosis during pregnancy: 3 couples do not need prenatal diagnosis during pregnancy.
Conclusion: deafness family re birth of deaf children with high probability, the re birth before genetic testing can predict fetal risk.

【學(xué)位授予單位】：揚(yáng)州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R764.43

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