Leber遗传性视神经病少见mt-DNA原发突变位点研究

目的探讨中国人Leber遗传性视神经病(LHON)患者少见线粒体DNA(mt-DNA)原发性突变位点。设计临床研究。研究对象常见线粒体DNA原发突变位点阴性的可疑LHON患者和正常人。方法采集临床诊断LHON患者的外周血,提取总DNA,聚合酶链反应(PCR)扩增和DNA直接测序的方法,进行mt-DNA G11778A、G3460A和T14484C突变位点筛查,对于以上3个突变位点均阴性的患者和正常对照组进行mt-DNA片段3962～4356和11320～11789扩增、测序,采用NCBI的BLAST服务,与剑桥标准序列进行比对,分析LHON患者是否携带C4171A或G11696A突变位点。主要指标mt-DNA测序结果。结果共收集临床诊断LHON患者56例,44例患者为3个常见位点之一为阳性(G11778A阳性36例,G3460A阳性3例,T14484C阳性5例),3个常见原发突变位点均为阴性患者12例,其中1例患者携带C4171A突变,未发现G11696A突变位点。正常对照组25例,未发现G11696A、C4171A突变位点。结论C4171A突变位点可能是中国人LHON患者少见原发突变位点。(眼科,2007,16:382-385)     

首诊为视神经炎的急性视力下降81例病因分析

目的：了解初诊为视神经炎的急性视力下降患者的病因。方法：收集临床初诊为视神经炎的急性视力下降患者81例，进行详细的临床检查以及相应的实验室、电生理学和影像学检查，综合分析病例重新诊断。结果：81例初诊为视神经炎的急性视力下降患者中，49例(60.5％)符合视神经炎临床诊断；其中23例符合炎性脱髓鞘性视神经炎的临床和实验室特点。9例患者的视神经炎以鼻窦炎为可能病因；少数病例为结核性、病毒性或梅毒性视神经炎；13例视神经炎患者病因不明，其中5例临床疑诊为Leber遗传性视神经病变(LHON)者3例进行线粒体DNA(mt-DNA)11778位点突变检查，2例阳性。81例患者中有32例不符合视神经炎的诊断，其中以缺血性视神经病变、心因性视力下降和颅内肿瘤等疾病造成的视神经压迫和／或视乳头水肿较常见。结论：缺血性视神经病变等很多疾病易与视神经炎混淆，正确诊断视神经炎需要严格和详细的临床和实验室检查；对于病因不明的视神经炎患者，进行与LHON相关的mt-DNA突变的筛查，可能有助于其诊断和鉴别诊断。     

孤发性Leber遗传性视神经病Wallace突变的检测

自Wallace等首次发现Leber遗传性视神经病(Leber′shereditaryopticneuropathy,LHON)与线粒体DNA(mt-DNA)np11778G→A突变密切相关[1],及其后np3460等点突变的发现以来,通过基因检测方法诊断LHON变为现实。我们对一无家族史、原因不明的视神经萎缩病例进行了上述点突变的检测,现报道如下。1　资料和方法1.1　临床资料患者男,6岁。因双眼视力下降半年就诊。曾在当地医院诊断为双眼弱视,后又诊断为视神经炎。曾作头部CT检查未见异常。患儿为独生子,否认家族中有相同病史。视力:右眼0.2,左眼0.3,均不能矫正。双眼屈光间质透明,视盘色…     

散发性视神经炎的线粒体DNA11778位点检测

Leber病系线粒体DNA基因突变引起的母系遗传性疾病,有家族史者易诊断。现将一组散发性视神经炎患者进行mt-DNA 11778位点突变检查,结果报告如下。     

我国睑裂狭小综合征患者FOXL2基因突变研究

目的 明确我国睑裂狭小综合征(BPES)患者Forkhead box L2 (FOXL2)基因突变位点及突变类型,分析突变引起的蛋白质结构变化,以提高疾病的诊断准确率.方法 实验研究.2008年1月至2009年12月在浙江大学医学院附属第二医院眼科中心收集到5例我国BPES患者,采集患者外周静脉血,提取基因组脱氧核糖核酸(DNA),应用聚合酶链反应和DNA测序技术检测FOXL2基因编码序列,并与患者家属及110名正常人相应序列进行比对.结果 两个家系中BPES患者均检测到c.672_701dup30(p.Ala224_Ala234dup)杂合突变,3例散发患者分别检测到c.655C＞T(p.Q219X)、c.370 A＞G(p.K124E)和c.858_874dup17(p.P292fs)杂合突变.结论 在BPES患者中发现FOXL2基因c.370A＞G(p.K124E)和c.858_874dup17(p.P292fs)两种新的杂合突变,扩展了国内外FOXL2基因突变谱,在两个家系中均发现c.672_701dup30(p.Ala224 _Ala234dup)杂合突变,再次证明这一位点为国内BPES患者FOXL2基因突变热点.     

Leber''''s遗传性视神经病变患者的线粒体DNA检测

目的　探讨与Leber's遗传性视神经病变 (Leber'shereditaryopticneuropathy ,LHON)相关的线粒体DNA原发位点突变在视神经疾病中的诊断意义。方法　 79例各种原因引起的双侧视神经疾病中 ,16例为临床诊断的LHON患者 ,44例为可疑LHON患者 ,2例为酒精性弱视患者 ,4例为多发性硬化症患者 ,5例为常染色体显性遗传的视神经萎缩患者 ,4例为原发性开角型青光眼患者 ,3例为脊髓小脑退行性变和 1例乙胺丁醇引起的视神经萎缩患者。用聚合酶链反应 (polymerasechainreaction ,PCR)及限制性片段长度多态性技术 ,检测外周血DNA中提取的线粒体DNA的 3 46 0位点、11778位点及 144 84位点 ,分析 3个原发位点的突变。结果　 31例 (39 2 % )呈 11778位点突变阳性 ,其中包括 16例临床诊断为LHON的患者、13例 (2 9 5 % )可疑LHON患者及 2例酒精性弱视患者。余 48例均未检出上述 3个原发位点突变。结论　线粒体DNA的检测分析为确立或排除LHON提供了诊断依据 ,尤其是对无家族史或原因不明的双侧性视神经炎的患者更具有诊断价值。     

Leber视神经萎缩分子生物学检测

目的 探讨线粒体DNA1778突变与Leber视神经萎缩(Leber shereditary optic neuropathy,LHON)之间的关系. 方法采用突变特异性引物PCR(mutation specific primer PCR)及PCR RFLP(MaeⅢ)检测临床可疑LHON患者及有关亲属，两种方法结果均阳性者则收集其临床资料。 结果 10例11778突变阳性者中1例为携带者，9例为LHON病人,患者中男性6例，女性3例，起病年龄12-25岁，双眼起病间隔0-6个月，随访0-12年,视力为指数/眼前-0.1(除一例生育小孩后发病的患者有视力恢复),接受过视野、视诱发电位和色觉检查的患者结果均异常，而视网膜电图检查结果及全身情况多正常。 结论 10例受试者的两种分子生物学检测结果都显示线粒体DNA11778突变。携带者状态和视力恢复现象的存在表明线粒体DNA突变虽是LHON的主要病因，但其他因素如内分泌失调也可影响其发病。(中华眼底病杂志,1998,14:156-158)     

扬州市278例非综合征性耳聋患者常见耳聋基因突变检测结果分析

目的调查扬州地区非综合征性耳聋(NSHI)患者分子流行病学特征,了解耳聋的分子病因。方法所选对象为江苏省扬州市中、重度NSHI患者,共278例。利用耳聋基因芯片诊断试剂盒和直接测序2种方法筛查4个国人中常见的耳聋相关基因中的9个热点突变,包括GJB2(35delG、176del16、235delC、299del AT),GJB3(538C>T),SLC26A4(IVS7-2A>G、2168A>G)和线粒体DNA 12S rRNA(A1555G及1494C>T)突变。结果在278例耳聋患者中,2种方法共检出145例携带致聋突变(突变比率达到52.2%)。GJB2突变患者人数88例,其中纯合突变51例(18.3%)、单杂合突变25例(9.0%)、复合杂合突变12例(4.3%)。SLC26A4纯合突变14例(5.0%)、单杂合突变26例(9.4%)、复合杂合13例(4.7%)。235delC纯合突变/SLC26A4 IVS7-2A>G杂合突变1例(0.4%),235delC纯合突变/SLC26A4 IVS7-2A>G杂合突变1例(0.4%),线粒体DNA12SrRNA A1555G突变2例(0.7%);阴性133例(47.8%)。结论扬州地区耳聋患者GJB2突变高于全国的遗传性耳聋平均发生率。应用耳聋基因诊断技术可以在耳聋患者病因调查中进行快速诊断筛查,值得推广应用。     

中国Leber遗传性视神经病变14484位点突变的家系分析

目的:了解我国Leber遗传性视神经病变(Leber’shereditaryopticneuropathy,LHON)线粒体DNA(mtDNA)14484位点突变患者的发病率和临床特征。方法:对来自117个家系的119例临床确诊或疑诊LHON的患者进行mtDNA检测。对3例证实为14484位点突变的家系做深入调查并收集相关病史及临床资料,抽取15例家属的血样进行mtDNA检测。结果:存在线粒体DNA突变的62例(62/119,52.1%)中,14484位点突变仅3例(4.8%)。该3例3个家系56例中,28例有眼部症状,外显率50%。发病经过和临床表现类似11778位点突变的LHON,但其中视力恢复者17例(60.7%)。15例家属的血样检测再次证实为14484位点突变。结论:我国LHON患者中14484位点突变者少见,其临床表现与11778位点突变者相似,惟视力恢复率高。     

眼睑黄色瘤与高胆固醇血症中遗传因素的相关性研究

目的:探讨眼睑黄色瘤(XP)与高胆固醇血症中遗传因素的相关性,为揭示其发病机制提供依据。方法:选取2019-11/2021-01在佛山市三水区人民医院眼科就诊的XP患者29例,抽取外周静脉血,通过二代测序(NGS)技术检测患者的基因突变情况,同时分析患者的血脂情况。结果:纳入的XP患者中有21例检出基因突变,其中13例为高胆固醇血症患者,8例为正常胆固醇患者,发生突变的基因包括STAP1、APOB、LDLRAP1、LDLR、PCSK9和APOE,基因突变类型包括3-UTR变异、框内缺失、错义变异、5-UTR变异、同义突变、内含子突变、可变剪接体变异、非编码转录外显子突变和非编码转录突变。结论:高胆固醇血症的遗传因素与XP疾病之间有一定的相关性。     

DNA diagnosis of Leber's hereditary optic neuropathy performed at Keio University Hospital

PURPOSE: To learn the clinical value of DNA diagnosis for Leber's hereditary optic neuropathy (LHON), we reviewed the results of DNA diagnosis performed at Keio University Hospital. METHODS AND PATIENTS: Included were 224 patients, 87 patients at Keio University Hospital and 137 patients from other clinics, with bilateral optic neuropathy who were suspected of having LHON. With informed consent, the 3460, 9804, 11,778, 13,730, and 14,484 mutations of mitochondria DNA (mt-DNA) were examined form 1990 to 1998. Percentage of male patients, age at onset of the disease, and percentage of familial history were compared between patients with and without the mutations. The clinical diagnosis at the time of DNA analysis were examined in patients without the mutation. RESULTS: Seventy two(32%) of the 224 patients had one of the five mtDNA mutations, 63(88%) patients had the 11,778 mutation, 6(8%) had the 14,484 mutation, and 3(4%) had the 3460 mutation. In 72 patients with one of the LHON mutations, 89% of the patients were male, the average age of the disease onset was 24.3 years, and 42% of the patients had a familial history of the disease. Eighty (53%) of 152 patients who did not have one of the 5 mutations were diagnosed as having bilateral optic atrophy with unknown causes. CONCLUSION: Although DNA diagnosis of LHON is a useful clinical test, we must know the clinical characteristics of the disease, before taking advantage of this analysis.     

12家系视网膜母细胞瘤基因诊断的回顾性分析

目的：检测并分析视网膜母细胞瘤患者的遗传学病因,为患者及其家系基因诊断、遗传咨询、产前诊断提供理论依据。方法：实验研究。收集12个家系视网膜母细胞瘤患者的临床检查资料,采集患者及家系受检者外周血提取基因组DNA,已故患者由家属提供石蜡切片组织提取基因组DNA,抽取相关孕妇羊水细胞提取基因组DNA。应用二代测序筛查、Sanger测序法验证RB1基因候选变异。通过相关数据库和PubMed文献检索相关变异的致病性,依据遗传变异分类标准指南判断并分级候选变异的致病性。依据家系先证者致病突变,相关孕妇行RB1基因产前诊断。结果：纳入的12个视网膜母细胞瘤家系,其中8个家系检测到致病基因变异,检测阳性率67%。检测到的8个不同RB1基因突变中,6个为已报道致病性变异;RB1基因c.2404dupG（p.N803Efs*12）和c.1380_1381insCTTA （p.K462Tfs*2）为本研究发现的新突变。变异致病性综合分析本研究检测到的8个不同RB1基因变异均为致病性。结论：本研究纳入的12个视网膜母细胞瘤家系中有8个家系患者检测到致病突变而获得基因诊断;本研究同时发现了导致视网膜母细胞瘤的新突变。相关家系依据基因诊断结果进行了有效的遗传咨询和产前诊断。     

Leber's hereditary optic neuropathy mutations in Korean patients with multiple sclerosis.

Several different mitochondrial DNA (mtDNA) sites for mutations of Leber's hereditary optic neuropathy (LHON) have been reported to be present in patients with multiple sclerosis (MS). To further study this association of LHON and MS in the Korean population, we tested 20 MS patients for the presence of mtDNA mutations at nucleotide (nt) 11778 in all 20 patients, and at nt 14484, nt 3460 and nt 15257 in 15, 12 and 12 patients, respectively. However, none of the MS patients exhibited any pathogenic LHON mtDNA mutations. In conclusion, we found no evidence for any association between MS and the LHON mutation in the Korean population.     

Phenotypic and genetic spectrum of Danish patients with ABCA4-related retinopathy

Background: Pathogenic variations in the ABCA4 gene were originally recognized as genetic background for the autosomal recessive disorders Stargardt disease and fundus flavimaculatus, but have expanded to embrace a diversity of retinal diseases, giving rise to the new diagnostic term, ABCA4-related retinopathy. Diagnostic genotyping of ABCA4 is complicated by the large size of the gene and the existence of approximately 600 known pathogenic variations, along with numerous rare polymorphisms. A commercial diagnostic array-based assay has been developed targeting known mutations, however a conclusive genetic diagnosis must rely on a comprehensive genetic screening as the mutation spectrum of ABCA4-related retinopathies continues to expand. Material and methods: Among 161 patients with a Stargardt-related phenotype previously assessed with the commercial ABCA4 mutation microarray, we analyzed the ABCA4 gene with High-resolution melting (HRM) in patients in whom the array analysis identified either a heterozygous mutation (n?=?50) or no mutation (n?=?30). Results: The HRM method detected each of the already known mutations and polymorphisms. We identified the second ABCA4 mutation in 31 of 50 heterozygous patients (62%). Several novel mutations were identified of which four were identified multiple times. The recurrent novel mutations were subsequently assessed among the 30 patients with possible ABCA4-related diseases, previously found to be negative for known ABCA4 mutations by array analysis. In total, 30 different mutations were identified of which 21 have not been described before. Conclusion: Scandinavian patients with ABCA4-related retinopathy appear to have a distinct mutation spectrum, which can be identified in patients of diverse clinical phenotypes.     

携带ND1基因G3635A突变的Leber遗传性视神经病变三家系线粒体基因突变位点检测及其分子遗传致病机制

目的 观察3个ND1基因G3635A突变Leber遗传性视神经病变(LHON)家系线粒体基因组中的突变位点,探讨其分子遗传致病机制。方法 3个家系共88名成员纳入本研究。母系成员53名,非母系成员35名。分别采用标准对数视力表、佳能眼底数码彩色照相、Humphrey视野计、俞自萍色觉图、德国罗兰电生理仪对所有成员行视力、眼底、视野、色觉及视觉诱发电位检查。其中,确诊为LHON 16例,未患LHON 72名。选择135名无血缘关系的中国温州地区正常健康者作为对照组。抽取所有受试者外周静脉血,提取全基因组DNA,检测ND1基因G3635A突变位点。采用扩增产物片段有重叠的24对引物,检测3个家系先证者线粒体单体型分型和基因组突变位点。结果 3个家系先证者及母系成员均发现ND1基因G3635A突变位点,非母系成员和对照组受试者均未发现ND1基因G3635A突变位点。先证者线粒体单体型分型分别为东亚单体型N9a3、D4、R11a。先证者线粒体全基因组检测发现,除ND1基因G3635A突变位点外,D-Loop区存在12个变异位点,RNA编码区存在6个变异位点,多肽编码区存在36个变异位点。结论 3个ND1基因G3635A突变家系先证者及母系成员均存在G3635A突变位点;G3635A突变位点是3个家系的分子遗传致病基础。     

Comprehensive screening of the USH2A gene in Usher syndrome type II and non-syndromic recessive retinitis pigmentosa

A screen of the entire coding region of the USH2A gene in 129 unrelated patients with Usher syndrome type II (USH2) and in 146 unrelated patients with non-syndromic autosomal recessive retinitis pigmentosa (ARRP) uncovered 54 different sequence variations, including 18 likely pathogenic mutations (13 frameshift, three nonsense, and two missense), 12 changes of uncertain pathogenicity (11 missense changes and one in-frame deletion), and 24 non-pathogenic rare variants or polymorphisms. Of the 18 likely pathogenic mutations, nine were novel. Among the USH2 patients, 50 (39%) had one or two likely pathogenic mutations. The most common mutant allele in USH2 patients was E767fs, which was found in 29 patients, including one homozygote. Among the ARRP patients, we found 17 (12%) with one or two likely pathogenic mutations. The most common mutant allele in ARRP patients was C759F and it was found in 10 patients. The C759F allele was also found in two USH2 patients; in neither of them was a change in the other allele found. The second most common mutant allele in both patient groups was L1447fs (found in 6/50 USH2 patients and 6/17 ARRP patients). Of the 50+17=67 patients with identified USH2A mutations, only one mutation in one allele was found in 41+12=53 (79%); the reason for the high proportion of patients with only one identified mutation is obscure. Our results indicate that USH2A mutations are found in about 7% of all cases of RP in North America, a frequency similar to the RPGR gene (8%) and the rhodopsin gene (10%).     

Leber遗传性视神经病变线粒体DNA突变观察

目的探讨国人Leber病线粒体DNA3个原发位点突变的发生频率及其相关研究。方法视力、视野、视觉电生理、FFA等检查而确诊为视神经病变者,常规采用外周血液行mtDNA3个位点检测,行单链构像多态分析、突变特异引物多聚酶反应、改良等位基因特异性PCR及序列分析等;同时对发病性别、年龄、视力等进行观察。结果65个家系中有104例mtDNA突变。11778位点阳性,发病97例,男73例,女24例;携带者41例,男8例,女33例。mtDNA14484位点突变9例,5例发病;3460位点突变3例,2例发病(同时并发11778位点突变)。其发生频率11778占93.3%,14484占4.8%,3460和11778占1.9%.发病年龄15岁以下占48%,25岁以下93.3%.视力低于0.05者占60.6%.长期随访中有10例视力不同程度恢复,自发恢复率占9.6%.结论Leber遗传性视神经病变以男性mtDNA11778位点突变占绝对多数,可作为国人常规初筛,属母系遗传,严重威胁视力,发病年龄有偏低趋势,视力恢复与不同位点有关。     

Leber’s hereditary optic neuropathy mitochondrial DNA mutations in familial multiple sclerosis

Leber’s hereditary optic neuropathy (LHON) can be difficult to distinguish from optic neuritis due to multiple sclerosis (MS). For several decades an association of LHON and MS has been suspected, and within the past 7 years the LHON nucleotide (nt)-3460 and nt-11778 mtDNA mutations have been identified in several patients with MS-like phenotypes. To further study this association, we tested 42 index patients with clinically definite, familial MS for the LHON mtDNA mutations at nt-3460, nt-11778, and nt-14484. No patients had a pathogenic LHON mtDNA mutation; however, two MS patients with unilateral optic neuritis harbored the nt-15257 mtDNA polymorphism that was reported originally as a pathogenic LHON mutation. Several investigators have shown evidence that the nt-15257 mtDNA mutation is not primarily pathogenic. Therefore, we conclude that pathogenic LHON mtDNA mutations are absent or rare in unselected patients with familial, clinically definite MS (95% confidence intervals for each of the negative mutations 0–7.0%). Received: 25 February 1998 Revised version received: 4 August 1998 Accepted: 20 August 1998     

原发性开角型青光眼MYOC- TIGR基因突变研究

目的 筛选并研究原发性开角型青光眼(POAG)小梁网糖皮质激素诱导反应蛋白MYOC- TIGR基因突变情况。方法 病例对照研究。抽取2002年1至12月就诊并诊断为POAG患者118例和150例非POAG对照者的外周静脉血4～8ml,采用酚-氯仿抽提全基因组DNA,应用聚合酶链反应(PCR)技术扩增全基因组DNA,以单核苷酸构象多态性(SSCP)分析MYOC基因的3个外显子（7对引物）编码区域序列改变。对SSCP分析异常者,采用双向测序法进一步证实。应用Cfr13I、Hinfl及BsmA1等限制性内切酶检测对照者MYOC基因编码区序列改变。POAG与非POAG人群MYOC基因各位点突变率比较采用x2检验。结果 基因序列分析发现G12R、I288M及Y353I共3个基因序列改变。118例POAG患者中仅有5例(4.23％)发生G12R位点突变,150例对照者中未见G12R位点改变;两组G12R位点突变率比较差异有统计学意义(x2=4.37,P=0.037)。POAG组和对照组均有I288M和Y353I位点改变,但两组突变率比较差异无统计学意义(x2=0.07,P=0.791和x2=0.56,P=0.453)。POAG患者基因突变率4.23％。结论 MYOC基因突变可能与POAG发病有关,MYOC基因突变可能是POAG发病的分子机制之一。