与耳聋相关的线粒体DNA A1555G突变的无创检测
Noninvasive Test for Mitochondrial DNA A1555G Mutation Associated with Deafness.
作者信息
Fan Wenlu, Zhu Yi, Tang Xiaowen, Xue Ling
出版信息
Clin Lab. 2017 Jan 1;63(1):127-131. doi: 10.7754/Clin.Lab.2016.160709.
BACKGROUND
The homoplasmic mitochondrial DNA (mtDNA) A1555G mutation in the highly conserved decoding site of 12S rRNA has been associated with aminoglycoside-induced and nonsyndromic hearing loss in many families worldwide. The A1555G mutation detection is an important part of hearing screening. At present, blood samples are the most common source of genomic DNA. However, drawing blood is invasive for individuals.
METHODS
The whole genomic DNA of samples carrying mitochondrial DNA A1555G mutation (LX010 and LX044) with aminoglycoside-induced and nonsyndromic deafness as well as wild type were used as templates. The PCR products were analyzed by DNA sequencing, PCR-RFLP, dot blot, and southern blot.
RESULTS
The DNA sequencing, PCR-RFLP, dot blot, and southern blot demonstrate that buccal cell DNA can be used for the screening and identification of the A1555G mutation as well as peripheral blood DNA.
CONCLUSIONS
This study established a convenient, noninvasive and suitable for clinical determination of mtDNA A1555G mutation associated with deafness.
背景
12S rRNA高度保守解码位点的同质性线粒体DNA（mtDNA）A1555G突变与全球许多家庭中氨基糖苷类药物诱发的非综合征性听力损失有关。A1555G突变检测是听力筛查的重要组成部分。目前，血液样本是基因组DNA最常见的来源。然而，采血对个体具有侵入性。
方法
以携带线粒体DNA A1555G突变（LX010和LX044）且患有氨基糖苷类药物诱发的非综合征性耳聋的样本以及野生型样本的全基因组DNA作为模板。通过DNA测序、PCR-RFLP、斑点印迹和Southern印迹分析PCR产物。
结果
DNA测序、PCR-RFLP、斑点印迹和Southern印迹表明，颊细胞DNA可用于A1555G突变的筛查和鉴定，外周血DNA也可用于此。
结论
本研究建立了一种方便、无创且适用于临床检测与耳聋相关的mtDNA A1555G突变的方法。
Zotero 插件