Liu Yalan, Sang Shushan, Ling Jie, He Chufeng, Mei Lingyun, Feng Yong
Department of Otolaryngology Head and Neck Surgery，Xiangya Hospital，Central South University，Changsha，410008，China.
Xiangya Hospital，Central South University，Province Key Laboratory of Otolaryngology Critical Diseases.
Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2020 Feb;34(2):153-157. doi: 10.13201/j.issn.1001-1781.2020.02.013.
To detect 20 common deafness gene mutations in non- syndromic deafness patients in China using PCR- RDB, and analyze and summarize the mutation data to explore the clinical value of this method. The PCR- RDB and Sanger sequencing were used to detect 20 common mutations of four deafness genes（, and ） in 500 patients with non- syndromic hearing loss . The Sanger sequencing was used to compare the sensitivity, specificity, positive predictive value, negative predictive value, and total coincidence rate of the deafness mutation detected by PCR- RDB. A total of 500 samples were detected. 147 wild- type samples, 81 homozygous mutant samples, 240 heterozygous mutant samples, 32 composite heterozygous mutant samples were detected using the PCR- RDB within the range of 20 gene mutations, which were identical to the Sanger sequencing results. GJB2 c.235delC and SLC26A4 c.919- 2 A>G are the most common hotspot mutations in this study, followed by mtDNA m. 1555 A>G. Compared with the Sanger sequencing method, the sensitivity, specificity, positive predictive value, negative predictive value, and total coincidence rate of the real- time fluorescence PCR melting curve method were 100%, and the Kappa value was one. PCR reverse dot-blot hybridization is a simple, rapid, sensitive and specific method for detecting 20 mutations of 4 common deafness genes in Chinese population, it is expected to be used in clinical detection of deafness genes in the future.
采用聚合酶链反应-反向点杂交（PCR-RDB）技术检测中国非综合征性耳聋患者20种常见耳聋基因突变情况，分析总结突变数据，探讨该方法的临床应用价值。应用PCR-RDB和桑格测序法对500例非综合征性听力损失患者的4个耳聋基因（、和）的20种常见突变进行检测。采用桑格测序法比较PCR-RDB检测耳聋突变的灵敏度、特异度、阳性预测值、阴性预测值及总符合率。共检测500份样本。在20个基因突变范围内，应用PCR-RDB检测出147份野生型样本、81份纯合突变样本、240份杂合突变样本、32份复合杂合突变样本，与桑格测序结果一致。GJB2基因c.235delC突变和SLC26A4基因c.919-2A>G突变是本研究中最常见的热点突变，其次是线粒体DNA m.1555A>G突变。与桑格测序法比较，实时荧光PCR熔解曲线法的灵敏度、特异度、阳性预测值、阴性预测值及总符合率均为100%，Kappa值为1。PCR反向点杂交技术是一种简便、快速、灵敏、特异的检测中国人4个常见耳聋基因20种突变的方法，有望用于今后耳聋基因的临床检测。
