口腔黏膜拭子在遗传性耳聋常见致病基因检测中的应用研究

Application of Buccal Swab in Detecting of Common Pathogenic Genes of Hereditary Deafness

目的：探讨口腔黏膜拭子在遗传性耳聋常见致病基因检测中的应用，为常见耳聋致病基因筛查提供无创的样本采集方式。方法：195例新生儿，经听力筛查被诊断为听力障碍，收集其口腔黏膜拭子和外周血样本，抽提DNA，核酸测定仪检测两种收集方式样本所得DNA质量。对两种样本进行三种常见遗传性耳聋致病基因GJB2基因、SLC26A4基因和线粒体基因12S rRNA全外显子巢式PCR扩增，比较两种方法扩增产物及测序效果。结果：口腔黏膜拭子抽提DNA质量较外周血低，差异具有统计学意义（P〈0．05）；在PCR扩增后，两种样本扩增产物凝胶电泳无明显差异，测序结果无明显差异（P〉0．05）。结论：口腔黏膜拭子作为一种无创的样本采集方式，可用于三种常见遗传性耳聋致病基因检测的样本采集。

Objective： To discuss the application of buccal swab in detection of three most common pathogenic genes of hereditary deafness and to provide non - invasive sample collection methods for the screening for common deafness pathogenic genes. Methods：Buccal swabs and peripheral blood samples of 195 newborns diagnosed as hearing impairment via universal newborn hearing screening process were collected. We extracted DNA of the samples obtained by the two methods of collection, determined the quality （purity and concentration） of the DNA. Among the DNA of samples, three common pathogenic genes of hereditary deafness （GJB2,SLC26A4, mitochondrial genes 12S rRNA） were amplified by using the nested polymerase chain reaction. We compared the amplified products and sequencing results of the DNA obtained by the two methods of collection. Results：There were statistical differences on quality of the DNA obtained by buccal swabs and peripheral blood （P = 0. 000, P 〈 0.05 ）. The quality of the DNA obtained by buccal swabs was inferior. After amplification of nested PCR, there were no significant statistical differences on the results of gel electrophoresis （ P = 0.315, P 〉 0.05 ）. The same result was obtained in sequencing. Conclusions： As a non - invasive sample collection, buccal swabs can be used for the detection of three common deafness pathogenic genes.