本文選題：leber遺傳性視神經(jīng)病　切入點(diǎn)：線粒體DNA　出處：《鄭州大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：背景 Leber遺傳性視神經(jīng)病變(LHON)是一種嚴(yán)重危害青少年視功能的線粒體眼病,臨床表現(xiàn)主要為雙眼中心視力急性或亞急性減退,一般不伴眼球疼痛或壓疼。視野檢查顯示中心暗點(diǎn)和旁中心暗點(diǎn)。LHON的流行病學(xué)調(diào)查顯示,其在全世界的發(fā)病率為1／31000,在荷蘭和芬蘭的發(fā)病率分別為1／39000和1／50000。目前公認(rèn)的LHON原發(fā)性致病位點(diǎn)有3個(gè),分別是位于ND4區(qū)的G11778A位點(diǎn),NDI區(qū)的G3460A位點(diǎn)和ND6區(qū)的T14484C位點(diǎn),95%的LHON由以上三個(gè)原發(fā)致病突變位點(diǎn)中的一個(gè)所致。在我國(guó)其突變率分別為92.9%、1.4%和5.7%,G11778A突變最為常見(jiàn),T14484C突變次之,G3460A突變罕見(jiàn),該病散發(fā)病例較高,起病及病程多樣化,在臨床上極易出現(xiàn)漏診或誤診,因此利用基因診斷篩查L(zhǎng)HON患者,可以快速準(zhǔn)確地篩查出突變位點(diǎn),對(duì)于LHON的預(yù)防和治療都有積極的意義。 目的 探討中原地區(qū)LHON患者的線粒體DNA(mtDNA)突變類型及遺傳特性,篩查中國(guó)人LHON患者新的繼發(fā)位點(diǎn)。 方法 對(duì)189例LHON疑似病例(包括3個(gè)家系)首先進(jìn)行眼科常規(guī)檢查和腦部MRI檢查,然后再分別應(yīng)用等位基因特異性PCR(MSP-PCR),聚合酶鏈反應(yīng)-限制性片段長(zhǎng)度多態(tài)性(PCR-RFLP)和聚合酶鏈反應(yīng)-單鏈構(gòu)象多態(tài)性(PCR-SSCP)聯(lián)合DNA測(cè)序的方法,對(duì)189例臨床癥狀疑診為L(zhǎng)HON患者的G11778A,G3460A和T14484C3個(gè)原發(fā)位點(diǎn)進(jìn)行mtDNA檢測(cè)。對(duì)于缺乏3個(gè)原發(fā)突變位點(diǎn)的疑似患者,進(jìn)行線粒體全基因組測(cè)序,尋找與疾病相關(guān)的其他繼發(fā)突變位點(diǎn)。 結(jié)果 1、在189例疑似LHON患者中,檢測(cè)出86例為原發(fā)性突變G11778A,G3460A和T14484C3個(gè)致病位點(diǎn)中的一個(gè)。 2、在86例原發(fā)性突變中,3460位點(diǎn)突變6例,占7%；11778位點(diǎn)突變66例,占77%；14484位點(diǎn)突變14例,占16%。 3、在被檢出的14例14484位點(diǎn)突變患者中,3例為單純性14484原發(fā)位點(diǎn)突變,占22.2%,均為男性；5例14484原發(fā)位點(diǎn)合并14502繼發(fā)位點(diǎn)突變,占33.3%,男1例,女2例；4例14484位點(diǎn)合并14470位點(diǎn)突變,占33.3%,男2例,女1例；2例14484位點(diǎn)合并14569位點(diǎn)突變,占11.1%。 4、通過(guò)對(duì)線粒體全基因組測(cè)序,發(fā)現(xiàn)A2706G,A1438G,T2115C,G3538T,G3834A,C4262A,C4275G,C7028T,G8392A,A10485G和C10719T這幾個(gè)突變位點(diǎn),在正常人當(dāng)中,也存在以上部分位點(diǎn)突變現(xiàn)象。 結(jié)論 1、中原地區(qū)LHON患者仍然以G11778A位點(diǎn)突變?yōu)橹?其次為T(mén)14484C,G3460A突變最少見(jiàn)。 2、T14484C位點(diǎn)突變者當(dāng)中,有單純性14484位點(diǎn)突變者,有14484合并14502位點(diǎn)突變者,有14484合并14569位點(diǎn)突變者,有14484合并14470位點(diǎn)突變者,ND6區(qū)是線粒體DNA突變研究的熱點(diǎn)之一。 3、通過(guò)對(duì)線粒體全基因組測(cè)序,發(fā)現(xiàn)一些新的繼發(fā)突變位點(diǎn)可能對(duì)原發(fā)位點(diǎn)的致病起協(xié)同作用,具體作用如何還有待于進(jìn)一步考證。
[Abstract]:Background. Leber's hereditary optic neuropathy (LHON) is a mitochondrial ophthalmopathy that seriously endangers the visual function of adolescents. The clinical manifestation is acute or subacute decrease of binocular central visual acuity. There is no pain or pressure in the eyeball. The visual field examination showed that the central dark spot and the paracentric dark spot. LHON epidemiological survey showed, It has an incidence of 1 / 31000 in the world, 1/39000 in the Netherlands and 1 / 50000in Finland. There are now three recognized primary pathogenic sites for LHON. G3460A of G11778A in ND4 region and 95% of T14484C locus of ND6 were caused by one of the three primary pathogenicity mutations. In China, the mutation rates of G11778A and G11778A were 92.91.4% and 5.775% respectively. The sporadic cases of the disease are high, the onset and the course of the disease are various, and it is easy to misdiagnose or miss diagnosis in clinic. Therefore, the mutation sites can be quickly and accurately screened by gene diagnosis in LHON patients. It has positive significance for the prevention and treatment of LHON. Purpose. To investigate the type and genetic characteristics of mitochondrial DNA mtDNA mutation in LHON patients in central China, and to screen new secondary sites in Chinese patients with LHON. Method. Routine ophthalmologic examination and brain MRI examination were performed on 189 suspected LHON patients (including 3 families). Then the methods of allele-specific PCR- MSP-PCRN, polymerase chain reaction-restriction fragment length polymorphism PCR-RFLP and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) combined with DNA sequencing were used, respectively. The G11778Agna G3460A and T14484C3 primary loci were detected by mtDNA in 189 patients with suspected clinical symptoms suspected to be LHON. For the suspected patients lacking three primary mutation sites, the whole mitochondrial genome was sequenced to find other secondary mutation sites associated with the disease. Results. 1. Of the 189 suspected LHON patients, 86 were found to be one of the primary mutations G11778Agna G3460A and T14484C3. (2) among 86 cases of primary mutation, there were 6 cases of mutation at No. 3460 locus, 66 cases of mutation at site 711778, 14 cases of mutation at locus 7714 484, accounting for 16000 cases. 3. Of the 14 patients with 14484 locus mutation, 3 were simple 14484 primary mutation (22. 2%), 5 male (14484) with 14502 secondary mutation (33. 3%), 1 male (1 male) and 2 females (4 / 4) with 14470 mutation at 14484 locus. 33.3%, male 2 cases, female 1 case with 14484 locus with 14569 locus mutation, accounting for 11.1%. 4. By sequencing the whole mitochondrial genome, we found that some of the mutation sites A2706G, A1438G, T2115CN, G3538TN, C4262Afen, C4275G, C7028T, G8392Agnon A10485G and C10719T, were also found to be mutated in normal subjects. Conclusion. 1. G11778A mutation was the main mutation in LHON patients in central China, followed by G3460A mutation in T14484Cad. Among the mutants with T14484C locus, there were simple 14484 mutation, 14484 with 14502 mutation, 14484 with 14569 mutation and 14484 with 14470 mutation. Nd6 region was one of the hotspots in mitochondrial DNA mutation research. 3. By sequencing the whole mitochondrial genome, we found that some new secondary mutation sites may have synergistic effect on the pathogenicity of the primary loci.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R774.6

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