Whole mitochondrial genome screening in maternally inherited non-syndromic hearing impairment using a microarray resequencing mitochondrial DNA chip
Whole mitochondrial genome screening in maternally inherited non-syndromic hearing impairment using a microarray resequencing mitochondrial DNA chip
复制标题
DOI：
10.1038/sj.ejhg.5201891
复制
发表时间：
2007-11-01
影响因子：
5.2
通讯作者：
Denoyelle, Francoise
中科院分区：
文献类型：
--
作者：
Leveque, Marianne;Marlin, Sandrine;Denoyelle, Francoise
线粒体DNA(MtDNA)突变被认为是非综合征性听力损失的主要或诱因。由于通常只有一部分线粒体基因组在耳聋中被研究，它的患病率可能被低估了。在通过法国合作网络收集的1350个非综合征性感音神经性耳聋家系中，我们选择了29个具有明确母系血统的大家庭，并对他们进行了12S rRNA、tRNASer(UCN)和tRNALeu(UUR)基因的mtDNA突变筛查。当没有发现突变时，使用微阵列重测序芯片：Affymetrix Inc.开发的MitoChip Version 2.0进行线粒体全基因组筛查。在29个家系中的9个中发现了已知的mtDNA突变，文中描述了这些突变：5个带有A1555G，2个带有T7511C，1个带有7472insC，1个带有A3243G突变。在剩下的20个家系中，有丝分裂芯片检测到258个线粒体同质变异和107个潜在的异质变异。通过对所选片段的直接测序进行对照，结果显示有丝分裂芯片的敏感性很高，但特异性很低，特别是对异质性变异。在物种保护、频率和系统发育调查的基础上进行了原始性分析，以筛选出更可能的致病变异。全基因组分析使我们能够确定另外五个具有推测致病线粒体变异的家族：T669C、C1537T、G8078A、G12236A和G15077A。这些结果表明，新的MitoChip平台是一种快速而有价值的工具，用于鉴定耳聋患者的新的mtDNA突变。
Mitochondrial DNA ( mtDNA) mutations have been implicated in non-syndromic hearing loss either as primary or as predisposing factors. As only a part of the mitochondrial genome is usually explored in deafness, its prevalence is probably under-estimated. Among 1350 families with non-syndromic sensorineural hearing loss collected through a French collaborative network, we selected 29 large families with a clear maternal lineage and screened them for known mtDNA mutations in 12S rRNA, tRNASer(UCN) and tRNALeu(UUR) genes. When no mutation could be identified, a whole mitochondrial genome screening was performed, using a microarray resequencing chip: the MitoChip version 2.0 developed by Affymetrix Inc. Known mtDNA mutations was found in nine of the 29 families, which are described in the article: five with A1555G, two with the T7511C, one with 7472insC and one with A3243G mutation. In the remaining 20 families, the resequencing Mitochip detected 258 mitochondrial homoplasmic variants and 107 potentially heteroplasmic variants. Controls were made by direct sequencing on selected fragments and showed a high sensibility of the MitoChip but a low specificity, especially for heteroplasmic variations. An original analysis on the basis of species conservation, frequency and phylogenetic investigation was performed to select the more probably pathogenic variants. The entire genome analysis allowed us to identify five additional families with a putatively pathogenic mitochondrial variant: T669C, C1537T, G8078A, G12236A and G15077A. These results indicate that the new MitoChip platform is a rapid and valuable tool for identification of new mtDNA mutations in deafness.