无锡地区新生儿耳聋基因的MALDI-TOF-MS筛查分析  

MALDI-TOF-MS based diagnosis and analysis of newborn hereditary deafness screening in Wuxi, Jiangsu Province

作　　者：孟西娜[1] 张婷[1] 臧嘉[1] 蒋新液[1] 陈钰[1] 侍佳[1] 吴志君[1] 许飞[1] 

机构地区：[1]南京医科大学附属无锡妇幼保健院检验科,214002

出　　处：《中华检验医学杂志》2015年第2期102-105,共4页Chinese Journal of Laboratory Medicine

基　　金：江苏省六大人才高峰项目（2012WS-097）;江苏省妇幼保健科研课题（F201235）;无锡市医管中心重大扶持项目（YGZF1105）

摘　　要：目的分析无锡地区耳聋致病基因突变热点及携带频率，建立无锡地区新生儿听力及耳聋基因联合筛查的临床推广标准。方法根据中国人群常见耳聋相关基因热点突变设计，利用MALDI—TOF-MS飞行时间质谱检测技术，以2013至2014年期间无锡市妇幼保健院2796例新生儿为研究对象，采集足跟血并提取基因组DNA，针对中国人群特点，进行4个基因（GJB2，GJB3，SLC26A4和12srRNA）20个突变位点的基因突变检测，结合OAE及AABR听力筛查，并对结果进行t检验分析。结果检出耳聋基因突变新生儿158例，携带率为5．65％，其中单位点突变152例（5．44％），复合突变6例（0．21％）。在阳性检出样本中，GJB2及SLC26A4基因突变占总阳性样本的88．61％（140／158）。阳性突变位点数最多的是GJB2基因235delC位点，共检出73例（46．20％），其次是SLC26A4基因IVs7—2A〉G位点（30／158，18．99％）。此次检测中还发现了3例纯合突变样本，均是12srRNA基因1555A〉G突变。通过耳聋基因筛查结合听力筛查，共确诊4例中度及重度耳聋新生儿。结论MALDI—TOF—MS对非综合征型耳聋患者的突变检出率较高，与传统常用基因检测方法相比，具有检测位点多、覆盖率高、高通量等特点，能够满足临床对常见耳聋基因检测的要求，为耳聋基因突变携带儿未来的婚育、用药及提早干预提供了积极有效的遗传指导意义。Objective To investigate the hot spots of genetic mutations that are responsible for hearing loss of newborns and the carrier frequencies of varies mutations, and also, to set up a clinical pattern for newborn hearing concurrent genetic screening in Wuxi. Methods A total of 2 796 newborns participated in this study were all born in Wuxi Maternity and Child Health Care Hospital during 2013-2014. Twenty hot mutation spots of four genes （i. e. GJB2, GLB3, SLC26A4, 12s rRNA） were chosen according to the latest epidemiologic study and were analyzed with matrix-assisted laser desorption ionization-time-of-flight mass spectrometry （MALDI-TOF-MS）. OAE and AABR methods were both used to help with the diagnosis of newborn hearing loss. Results 158 of 2 796 newborns tested （5.65%） were positive with at least one hot mutation spot. The number of positively tested single site mutation were 152 （5.44%） , while the number of positively tested compound heterozygous mutations were 6 （0.21%）. The positive mutation sites in GJB2 and SLC26A4 ranked 88.61% among all the positive mutation sites tested. Mutation site 235delc within GJB2 gene were hottest positive site with a number of 73 newborns, followed by IVS7-2A 〉 G within SLC26A4 （ 18.99% , 30/158）. Three 1555A 〉 G homozygous mutations were also found in 12s rRNA. Four deafness newborns were found with newborn hearing concurrent genetic screening. Conclusions In comparison to the conventional detection methods, there could be more detection sites, greater coverage , and high throughput in MALDI-TOF-MS, and it would provid valuable guidance for newborns in the future.

关 键 词：听觉丧失 连接蛋白类 膜转运蛋白质类 突变 光谱法 质量 基质辅助激 光解吸电离 新生儿筛查 

分 类 号：R764.43[医药卫生—耳鼻咽喉科] R440[医药卫生—临床医学]