52例听力损失儿童及其父母耳聋基因芯片筛查结果分析

目的：对非综合征性先天性重度及以上感音神经性听力损失儿童及其父母进行耳聋相关基因检测,探讨耳聋基因芯片筛查在临床中应用的有效性和可行性。方法选择来自医院听力检测中心的47个听障儿童家庭,包括52例非综合征性先天性感音神经性听力损失患儿及其父母,应用遗传学耳聋基因芯片对47个家庭进行GJB2、GJB3、SLC26A4、线粒体12S rRNA4个常见耳聋基因9个检测位点的基因检测。结果146例受检者中,17个家庭的43例筛查结果阳性,其中16例听力损失患儿筛查阳性,筛查阳性率为30.8%。GJB2基因235delC位点纯合突变8例,GJB2基因235delC位点杂合突变20例,GJB2基因235delC位点和SLC26A4基因IVS7-2A〉G位点杂合突变1例,SLC26A4基因IVS7-2A〉G位点纯合突变2例,SLC26A4基因IVS7-2A〉G位点杂合突变10例,SLC26A4基因2168A〉G位点杂合突变2例。结论应用耳聋基因芯片检测技术能快速、高效地检测非综合征性耳聋患者的遗传性致病基因,适用于大规模群体耳聋基因的筛查,有助于临床医生从病因学角度辅助耳聋诊断,引入正确的康复干预措施,并为具有聋病易感基因的听力损失儿童家庭提供针对性的遗传咨询指导。     

遗传性耳聋基因芯片检测及其临床意义

目的：探讨遗传性耳聋基因芯片检测的临床意义。方法：对贵阳市42名非综合征性聋哑儿童及其父母进行耳聋病因问卷调查、纯音听阈测试或听性脑干反应测试,并采用耳聋基因芯片进行突变检测,对芯片检测结果为杂合突变的样本进一步行测序验证。结果：42名患儿中,7例（16．67％）存在GJB2235delC纯合突变,4例（9．52％）存在GJB2235delC杂合突变,1例（2．38％）存在GJB2235delC／299delAT复合突变,2例（4．76％）存在PDSIVS7—2A〉G纯合突变,2例（4．76％）存在PDSIVS7—2A〉G杂合突变,在分子水平明确诊断者占38．10％。结论：贵阳地区耳聋患者存在较高的遗传性耳聋发生率,耳聋基因芯片诊断技术可以应用在临床中进行快速筛查、诊断,并可达到防止再出生聋儿,指导聋儿康复等积极效果。     

耳聋基因芯片技术在耳聋病因诊断中的应用

目的使用耳聋基因芯片技术对耳科门诊耳聋患者进行病因诊断。方法收集26例明确为感音神经性听力下降的聋病患者,使用基因芯片检测试剂盒进行检测。结果 26例患者中先天性耳聋者10例,检出率为40.00%;成年感音神经性耳聋者13例,检出率为23.08%;突发性耳聋者3例,检出率为66.67%。结论遗传性耳聋基因检测试剂盒对于明确聋病患者致聋原因有一定的帮助,具有临床推广价值。     

儿童听力筛查未通过的原因及耳聋基因检测分析

目的探讨儿童听力筛查未通过原因及耳聋基因筛查的意义。方法扬州市就诊我院门诊的14岁及以下入学儿童听力筛查未通过245人,所有儿童均行声导抗及耳声发射检查,5周岁及以上儿童进行纯音测听检查,5周岁以下儿童进行行为测听检查,不能配合者行ABR检查。基因筛查采用4个基因(GJB2、SLC26A4、12SrRNA和GJB3)20个位点的高通量测序。结果复测通过64人(26.12%),耵聍栓塞及外耳道疾病31人(12.65%),分泌性中耳炎132人(53.88%),急慢性中耳炎12人(4.90%),先天性中耳胆脂瘤2人(0.82%),小儿听神经病1人(0.41%),感音神经性聋1人(0.41%),大前庭水管综合征2人(0.82%)。GJB2基因杂合突变9人(3.67%),未筛查出纯合突变或复合杂合突变;SLC26A4基因杂合突变1人(0.41%),纯合突变1人(0.41%);线粒体12SrRNA基因突变携带1人(0.41%),未发现GJB3基因突变携带者。结论听力和耳聋易感基因联合筛查对发现耳聋和迟发性耳聋具有协同作用,解除病因和给予正确指导可降低耳聋的发生率。     

新生儿遗传性耳聋及听力筛查

耳聋是人类一种最常见的感觉系统缺陷。在世界范围内新生儿中听力障碍率为0.1～0.3%,其中约50%系遗传因素所致。遗传性听力损失根据是否伴有耳外组织的异常或病变分为综合症性听力损失     

东莞户籍33810例新生儿听力筛查联合耳聋基因检测与分析

目的了解分析东莞地区新生儿常见耳聋基因的突变类型和突变携带率,并对听力筛查和耳聋基因联合检测进行评价。方法对2016年1月至2017年2月在东莞辖区内出生的33810例户籍新生儿同时进行听力筛查和耳聋基因检测,听力筛查包括耳声发射和听性脑干反应,耳聋基因检测采用耳聋基因芯片(微阵列芯片法)对我国常见的致聋基因GJB2(c.235del C,c.299_300del AT,c.176_191del16,c.35del G)、SLC26A4(IVS7-2A>G,c.2168A>G)、线粒体12Sr RNA(m.1555A>G,m.1494C>T)和GJB3(c.538C>T)进行检测。结果 33810例新生儿共检出1145个等位基因突变,突变携带率约为3.39%。其中GJB2突变661个,突变携带率约1.96%;SLC26A4突变364个,突变携带率约1.08%;线粒体DNA12Sr RNA突变74个,突变携带率约0.22%;GJB3突变46个,突变携带率约0.14%。在6242例新生儿听力筛查结果中,听力筛查未通过103例,未通过率1.65%。结论 c.235del C、IVS7-2A>G是东莞地区新生儿耳聋基因突变的主要类型。开展新生儿听力筛查与耳聋基因联合检测有助于了解本地区耳聋基因携带情况及患儿听力状况,提早发现先天性耳聋患者,高度预警药物性耳聋和迟发性耳聋,做到早发现、早诊断、早干预、早治疗,对降低耳聋发生率有重要意义。     

烟台地区3785例新生儿遗传性耳聋基因位点筛查分析

目的 评估烟台地区新生儿耳聋基因的突变情况,为遗传性耳聋患者的临床治疗提供参考。方法 采集3785例出生3d的新生儿足底血血斑,提取人基因组DNA。利用微阵列芯片杂交法检测4个遗传性耳聋基因(GJB2、GJB3、SLC26A4及mtDNA 12S rRNA)15个位点,并用Sanger测序法对突变位点进行确证。将烟台地区新生儿耳聋基因突变情况与山东省平均突变情况进行比较。同时,针对相同的检测方法(微阵列芯片检测法)和相同的筛查位点(4个基因15个位点),将烟台地区与淄博地区进行耳聋基因突变情况的比较。结果 3785例新生儿共检出255例耳聋基因突变携带者,突变率为6.74%(255/3785)。单基因突变242例,包括134例GJB2、85例SLC26A4、12例GJB3和11例mt DNA 12S rRNA,其突变率分别是3.55%、2.25%、0.32%和0.29%。两个位点复合突变13例,其中非等位基因复合突变6例,等位基因复合突变5例,2例核基因与线粒体基因复合突变。烟台地区耳聋基因总突变率和GJB2基因突变率明显高于山东省平均突变情况(χ²=32.20,...     

基因芯片法检测21例耳聋患者基因突变的研究

目的探讨基因芯片在耳聋基因筛查中的应用价值。方法对扬州市聋哑学校21例8～18岁的耳聋患者中4个耳聋相关基因上的9个热点突变进行检测,包括GJB2(35 delG、176 del16、235 delC及299 delAT)、GJB3(C538 T)、SLC26 A4(IVS7-2 A>G、A2168 G)以及线粒体12 S rRNA(A1555G、C1494T)。结果 SLC26A4突变阳性率为38%(8/21),其中IVS7-2A>G杂合突变7例,IVS 7-2 A>G与A 2 1 6 8 G杂合突变1例;GJB 2突变阳性率为1 9%(4/2 1),其中1 7 6 del 1 6杂合突变1例,299 delAT杂合突变1例,176 del 16与235 del C杂合突变1例,235 del C纯合突变1例。结论基因芯片是一种筛查耳聋基因的高效、经济、简便、灵敏及特异性方法。     

湖南地区4132例备孕女性遗传性耳聋基因筛查结果分析

目的 探讨湖南地区4132例备孕女性遗传性耳聋基因筛查结果特点.方法 通过多重PCR扩增、高通量测序等技术联合,对18个耳聋致病基因的108个突变位点进行筛查,对阳性样本中的突变基因、位点以及突变阳性率等特点进行统计分析.结果 4132例检测样本中,阳性202例,检出率为4.89％.单基因突变型200例,包括SLC26...     

基因芯片技术及其在耳聋研究中的应用

基因芯片又称DNA芯片（DNAchip）、DNA微阵列（DNAmicroarray）,寡核苷酸阵列《oligonucleotjdearray）等。基因芯片技术是融微电子学．生物学、物理学、化学、计算机科学等多学科为一体的新技术。它可以将生物中许多不连续的分析过程．移植到固相的介质芯片,并使其连续化和微型化。随着基因芯片技术的逐渐成熟．基因芯片已逐步从实验室走向工业化,在生物医学领域获得了广泛应用。     

Molecular study of hearing loss in Minas Gerais,Brazil

IntroductionDeafness is the most frequent sensory deficit in humans. Incidence is estimated at 4:1000 births in Brazil. Specific programs for clinical care of patients with hearing loss are still scarce in Brazil and the issue is an important public health problem.ObjectiveTo determine the frequency of 35delG and D13S1830 mutations in GJB2 and GJB6 genes respectively in patients with non-syndromic sensorineural hearing loss from Minas Gerais, Brazil.MethodsThis research involved 53 individuals, who were assessed by a questionnaire for predicting the possibility of non-syndromic deafness and for data collecting. Samples were tested for the presence of the 35delG mutation in GJB2 gene and D13S1830 in GJB6 gene by polymerase chain reaction and restriction enzyme digestion.ResultsEpidemiological research has shown that the majority of the subjects are unaware of the etiology and the pathogenesis of hearing loss. In 9 patients (16.98%), 35delG mutation was found in heterozygosis and the allele frequency was estimate to be around 8.5%. Although 9.61% of the patients reported having some degree of consanguinity between the parents and 12.08% reported other cases of deafness in their families, this mutation was not found in homozygosis. The D13S1830 mutation was not found in this study.ConclusionThis research describes for the first time the frequency of the 35delG and D13S1830 mutation in hearing-impaired individuals from Minas Gerais, Brazil, and the collected data reinforce the need for further studies in this population due to heterogeneity of hearing loss.     

Absence of GJB6 mutations in Indian patients with non-syndromic hearing loss

Objective

Hearing loss is the most frequent sensory defect in human being. Genetic factors account for at least half of all cases of profound congenital deafness. The 13q11-q12 region contains the GJB2 and GJB6 genes, which code connexin 26 (CX26) and connexin 30 (CX30) proteins, respectively. Mutations in the gene GJB2, encoding the gap junction protein connexin 26, are considered to be responsible for up to 50% of familial cases of autosomal recessive non-syndromic hearing loss and for up to 15-30% of the sporadic cases. It has also been reported that mutations in the GJB6 gene contribute to autosomal recessive and autosomal dominant hearing defects in many populations. The 342-kb deletion [del(GJB6-D13S1830)] of the Cx30 gene is the second most common connexin mutation after the CX26 mutations in some NSHL populations. The aim of this study was to screen GJB6 gene mutations in Asian Indian patients with autosomal non-syndromic hearing loss.

Methods

We screened 203 non-syndromic hearing loss patients, who were negative for homozygous mutations in GJB2 gene, for GJB6-D13S1830 deletion and mutations in coding regions of GJB6 using polymerase chain reaction, denaturing high performance liquid chromatography and direct sequencing.

Results

No deleterious mutation in GJB6 gene was detected in our study cohort.

Conclusion

The present data demonstrated that mutations in the GJB6 gene are unlikely to be a major cause of non-syndromic deafness in Asian Indians.     

散发感音神经性聋患者中GJB2基因突变检测的临床意义

目的探讨散发感音神经性聋患者中GJB2基因突变检测的临床指导意义.方法运用聚合酶链反应对解放军总医院听力诊断中心收集的242例散发感音神经性聋患者(135例语前聋患者,107例语后聋患者)的GJB2基因编码区进行扩增,扩增产物纯化后直接测序分析.结果 135例语前聋患者中GJB2基因致病突变的复合杂合和纯合个体有26例,占语前聋个体的19.26%;107例语后聋患者中未发现复合杂合和纯合致病突变,仅发现3例235delC杂合突变携带者、1例176del16杂合突变携带者.结论语前聋者GJB2基因致病突变阳性率明显高于语后聋患者,语前聋患者常规进行GJB2基因检测可从基因水平明确诊断,并为耳聋患者提供重要遗传信息.     

Newborn hearing concurrent genetic screening for hearing impairment—A clinical practice in 58,397 neonates in Tianjin,China

Objective

Newborn hearing screening (NHS) is used worldwide due to its feasibility and cost-efficiency. However, neonates with late-onset and progressive hearing impairment will be missed by NHS. Genetic factors account for an estimated 60% of congenital profound hearing loss. Our previous cohort studies were carried out in an innovative mode, i.e. hearing concurrent genetic screening, in newborns to improve the abilities or early diagnosis and intervention for the hearing defects. In this study, we performed the first clinical practice of this mode in Tianjin city.

Methods

A large cohort of 58,397 neonates, born between December 2011 and December 2012, in 44 hospitals in Tianjin, were screened for 20 hot spot hearing loss associated mutations from GJB2, GJB3, SLC26A4 and MTRNR1(12S rRNA). The data of genetic screening results was comprehensively analyzed with newborn hearing screening (NHS) results.

Results

We developed an accurate, high throughput genetic screening method and applied it to a total of 58,397 newborns in Tianjin. 3225 (5.52%) infants were detected to carry at least one mutation allele in GJB2, GJB3, SLC26A4 or MTRNR1. 34 (0.58‰) infants were positive for hearing loss caused by GJB2 or SLC26A4 mutations (homozygote or compound heterozygote). 54(0.93‰) infants are heterozygous of various genes. 109(1.87‰) infants had the pathological mitochondrial DNA mutation.

Conclusion

Accurate, comprehensive hearing loss associated genetic screening can facilitate genetic counseling and provides valuable prognostic information to affected infants. This united screening mode of this study was a promising clinical practice.     

山西太原129例重度非综合征型感音神经性耳聋基因的筛查

目的应用耳聋基因芯片对重度极重度非综合征型感音神经性耳聋患者进行筛查。方法采集本地区聋哑学校和门诊散发的重度极重度非综合征型感音神经性耳聋患者129人的外周血并提取DNA,应用耳聋基因芯片检测GJB2,GJB3,SLC26A4,线粒体DNA(mitochondrial,mtDNA)12SrRNA热点突变位点。结果该耳聋人群中与筛查位点有关的耳聋比例占41.09%,共检出GJB2基因突变26例(20.16%);mtDNA突变8例(6.2%);SLC26A4基因突变21例(16.28%);未检出GJB3基因突变。结论本组耳聋人群中与筛查位点有关的耳聋比例高达41.09%,GJB2突变是该人群遗传性聋的最常见病因,SLC26A4突变为第二常见病因。     

湖北地区306例极重度聋患儿基因芯片筛查分析

目的：通过检测湖北地区极重度感音神经性聋患儿常见耳聋基因突变情况,分析该人群的分子病因学特点,为临床耳聋防治和遗传咨询提供参考。方法：收集306例湖北地区极重度感音神经性聋患儿,抽取外周血,提取DNA,应用遗传性耳聋基因芯片检测GJB2、GJB3、SLC26A4和线粒体12SrRNA4个基因的9个突变热点。对所有携带SLC26A4基因突变患者进行颞骨CT扫描。结果：306名患儿中,132例（43．14％）检出携带不同基因突变,其中有2例携带双基因突变。GJB2基因突变检出率为29．41％（90／306）,SLC26A4基因突变检出率为13．72％（42／306）,线粒体12SrRNA基因突变检出率为O．65％（2／306）。本组患者未检出GJB3基因突变。36例携带SLC26A4基因突变者颞骨CT扫描显示前庭水管扩大。结论：GJB2基因和SLC26A4基因是本组患儿最主要的致聋基因,其中235delC突变为最常见的突变位点,其次为1VS7—2A〉G突变。筛查SLC26A4基因常见突变有助于大前庭水管综合征的诊断。     

Mutational analysis for GJB2, GJB6, and GJB3 genes in Campania within a universal neonatal hearing screening programme

Abstract

Objective: To determine the incidence of GJB2 and GJB3 mutations and of two deletions upstream of the GJB6 gene in infants of the Campania region of southern Italy. Design: DNA samples from non-syndromic hearing-impaired infants enrolled in a neonatal screening programme for sensorineural hearing loss were analysed by PCR and by direct sequencing. The audiological features of infants with biallelic GJB2 mutations were also examined to identify genotype-phenotype correlations. Study sample: Molecular analyses were carried out in 129 affected and five unaffected infants. Results: A genetic etiology of hearing loss was identified in 28% of infants, including several at environmental risk of hearing loss. Neither GJB6 nor GJB3 (a gene not previously investigated in the Campania population) mutations were found. Conclusions: This study confirms the importance of universal neonatal hearing screening. The identification of a genetic cause in infants at environmental risk indicates that such infants should be included when investigating etiology. We confirm that also in our geographical area, c.35delG homozygotes tend to have severe symmetrical hearing loss, whereas hearing impairment is milder in compound heterozygotes.

Sumario

Objetivo: Determinar la incidencia de mutaciones GJB2 y GJB3 y de dos deleciones corriente arriba en el gen GJB6 en niñosde la región de Campania en el sur de Italia. Diseño: Se analizaron por PCR y por secuenciación directa muestras de ADN de niños con trastornos auditivos no sindrómicos incluidos en el programa de tamiz neonatal para hipoacusias sensorineurales. Los rasgos audiológicos de niños con mutaciones bi-alélicas del GJB2 también se examinaron para identificar correlaciones genotípicas y fenotípicas. Muestra del Estudio: Se realizó un análisis molecular en 129 niños afectados y en 5 no afectados. Resultados: Se identificó una etiología genética de la sordera en 28% de los infantes, incluyendo varios con riesgo ambiental de hipoacusia. No se encontraron mutaciones en el gen GJB6 o en el GJB3 (un gene no investigado previamente en la población de Campania). Conclusiones: Este estudio confirma la importancia del tamiz auditivo neonatal universal. La identificación de una causa genética en niños con un riesgo ambiental indica que tales infantes deberían incluirse cuando se investiga la etiología. Confirmamos que también en nuestra área geográfica, los homocigotos c.35delG tienden a tener hipoacusias simétricas severas, mientras que el trastorno auditivo es más leve en heterocigotos compuestos.     

GJB2 and mitochondrial DNA 1555A>G mutations in students with hearing loss in the Hubei Province of China

Objectives

The GJB2 and MTRNR1 1555A > G mutations are the prevalent causes of hearing loss worldwide. However, the mutation profiles of the two genes are dependent on the ethnic or geographic origins. Therefore, this study was to characterize the forms and frequencies of the two genes in 813 students with hearing loss in Hubei province, Central China.

Methods

Blood samples from 813 students were obtained with informed consent. Genomic DNA was extracted from peripheral blood leukocytes. The target fragments were amplified by polymerase chain reaction (PCR). Sequencing (or enzyme digestion) was applied to identify sequence variations.

Results

Ten different mutations were identified in GJB2 in 146 of the 813 (17.96%) patients and 11.81% (96/813) patients had homoplasmic mtDNA 1555A > G mutation.

Conclusions

This study demonstrated the high prevalence of GJB2 and mtDNA 1555A > G mutations in Central Chinese population. Therefore, it will be effective to perform GJB2 and mtDNA 1555A > G mutation analysis for genetic screening for hearing loss in this population.     

Functional hearing loss in children who were not aware of their hearing loss

We investigated one hundred and fourteen ears of 60 children (8 males, 52 females, aged from 6 to 13 years) with diagnoses of functional hearing loss (FHL), and were not aware of their own hearing loss. Forty nine (81.7%) of 60 cases examined were detected by school screening tests, 6 (10.0%) were referred to our hospital because their families noticed poor hearing responses, and 5 (8.3%) were enrolled because they complained of otalgia or discomfort in the ear. Forty (66.7%) showed only pure tone threshold loss without complications, and the remaining 20 associated nonorganic disorders. In addition, our investigation found 11 cases (18.3%) with nonorganic otalgia, 5 (8.3%) with functional visual disturbance, 1 (1.7%) with enuresis nocturna who refused to attend school, 1 with tinnitus, 1 with vertigo, and 1 with tic. Moreover, 11 (18.3%) of the 60 cases were suspected of being in conflict with school and/or home. The Type V Békésy pattern, which is frequently observed in FHL and it has clinical utility to distinguish FHL from other types of organic hearing loss, was detected in 44 ears (38.6%). Fifty two (45.6%) of 114 ears showed normal pure-tone thresholds during the clinical course. Sixteen (14.0%) ears needed more than 1 year for thresholds to normalize. These findings suggest that some FHL cases without awareness of their hearing loss resemble psychogenic hearing loss. In such cases, otolaryngologists should carefully check the patient's individual circumstances, and when appropriate, refer patients for psychiatric consultation.     

儿童双耳感音神经性聋临床特点及GJB2与GJB3基因突变分析

目的 探究并分析儿童双耳感音神经性聋临床表现特点及缝隙连接蛋白26（GJB2）、缝隙连接蛋白31（GJB3）基因突变情况。方法 本研究选取2020年3月—2021年3月接受治疗的150例双耳感音神经性聋患儿作为研究对象,纳入研究组;选择同期参与体检的150例听力正常的儿童作为对照组。试剂盒法提取血液白细胞中的基因组DNA,GJB2、GJB3基因采用编码区聚合酶链反应（PCR）进行检测。观察两组儿童GJB2和GJB3基因检测结果及研究组儿童耳聋程度、不同程度双耳感音神经性聋儿童的GJB2和GJB3耳聋基因突变率及基因突变位点情况、研究组儿童双亲GJB2、GJB3基因突变情况。结果 研究组儿童GJB2基因突变共50例（33.33%）,基因致病突变5种（35insG、95G>A、176-191del16、235delC、257C>G）,40例与235delC突变相关,占比80.00%;多态性基因改变3种（79G>A、341A>G、427C>T）;GJB3基因突变共6例（538C→T、547G→A）。对照组儿童GJB2基因突变共计3例（2.00%）,基因致病突变有两种（95G>A、235delC）;多态性基因改变有两种（79G>A、341A>G）;未发现GJB3基因突变。研究组儿童235delC突变率显著高于对照组（P<0.05）;研究组儿童中重度及极重度患者占达到59.33%（89/150）;不同程度双耳感音神经性聋儿童GJB2、GJB3基因突变率中极重度阳性检出率较高,分别为33.33%和7.5%;重度和危重度患儿致病突变位点例数显著高于轻中度患儿,多态性改变例数低于轻中度患儿（P均<0.05）。研究组儿童父母中发现GJB2基因突变父亲17例（5.67%）、母亲25例（8.33%）;GJB3基因突变父亲3例（1.00%）,母亲1例（0.33%）;父母亲均未检验出纯合突变和复合杂交突变以及GJB3基因突变。结论 目前临床暂无对双耳感音神经性聋的有效预防和诊断手段,而GJB2、GJB3基因突变检测可以作为产妇产前诊断的一种方法,从而对下一代进行预防,利于有效诊断耳聋的发生率,从而做到尽早预防,降低发病率。