Rapid identification of aminoglycoside-induced deafness gene mutations using multiplex real-time polymerase chain reaction
Rapid identification of aminoglycoside-induced deafness gene mutations using multiplex real-time polymerase chain reaction
复制标题
使用多重实时聚合酶链反应快速鉴定氨基糖苷类诱发的耳聋基因突变
DOI：
10.1016/j.ijporl.2015.04.028
复制
发表时间：
2015-07-01
影响因子：
1.5
通讯作者：
Dai, Pu
中科院分区：
文献类型：
--
作者：
Huang, Shasha;Xiang, Guangxin;Dai, Pu
背景资料：暴露于氨基糖苷类抗生素可在携带某些线粒体DNA（mtDNA）突变（C1494T和A1555G）的遗传易感个体中诱导耳毒性，导致听力损失。因此，需要一种快速的诊断方法来准确地识别携带此类基因mutation.Methods：在本研究中，我们描述了一种快速，可靠的四色，实时定量聚合酶链反应（qPCR）检测同时检测两个mtDNA 12S rRNA基因变异，A1555G和C1494T，这是在中国汉族人群中流行。结果：C1494T、A1555G和野生型DNA检测具有较高的灵敏度、特异性、重复性和准确性，可用于临床血液样本的基因分型。结论：这项研究表明，这种简单而廉价的方法可用于常规的分子诊断和潜在的大规模遗传筛查。(C)由Elsevier爱尔兰有限公司出版。
Background: Exposure to aminoglycoside antibiotics can induce ototoxicity in genetically susceptible individuals carrying certain mitochondrial DNA (mtDNA) mutations (C1494T and A1555G), resulting in hearing loss. So, a rapid diagnostic approach is needed to accurately identify subjects carrying such gene mutations.Methods: In the present study, we describe a rapid and reliable four-color, real-time quantitative polymerase chain reaction (qPCR) assay for simultaneously detecting two mtDNA 12S rRNA gene variants, A1555G and C1494T, which are prevalent in the Han Chinese population. This multiplex assay incorporates three allele-specific TaqMan probes labeled with different fluorophores in a single reaction, providing high genotyping accuracy for clinical blood samples.Results: Tests with C1494T, A1555G and wild-type DNA exhibited high sensitivity, specificity, reproducibility and accuracy of discriminating mutations from wild-type.Conclusions: This study shows that this simple and inexpensive method can be used for routine molecular diagnostics and potentially for large-scale genetic screening. (C) 2015 Published by Elsevier Ireland Ltd.