遗传性非综合征性常见耳聋基因诊断的研究

The study of diagnosis with the commom gene of hereditary nonsyndromic hearing loss.

目的应用三种方法对非综合征性感音神经性耳聋患者及其家属进行分子病因学研究,以期获得一种快速、简便、有效的临床诊断方法。方法采用酶切法、芯片法和测序法对国内报道突变频率比较高的6个耳聋基因进行检测。结果 19例耳聋患者,酶切法共检出4例GJB2 235delC,1例mt 12S rRNA;; A;;1555G;其中8例患者及1例患者的父母基因芯片法检出1例mt 12S rRNA;; A;;1555G,3例GJB2 235delC,3例SLC26A;;4基因突变,3例9位点均正常。mt 12S rRNA;;A;;1555G和GJB2 235delC基因芯片检测结果与酶切相一致。10例基因芯片检测及GJB3测序分析均未检出GJB3 538C>T突变及GJB3其他致病突变;GJB6基因序列未发现致病突变位点。mt 12S rRNA;; A;;1555G和GJB2 235delC位点测序分析结果与基因芯片和酶切结果相一致。SLC26A;;4基因突变IVS7-2A;;>G和2168A;;>G测序分析结果与基因芯片结果相符。随后对部分患者父母采用上述方法筛查,48例样品中,共检出15例携带致聋突变,检出率31.25%。结论酶切法、基因芯片法及测序法三者检查结果相符合。临床诊断因患者而异,综合三种检测技术,设计最佳筛查方式。

Objective： In order to searching a rapid, simple and effective genetic diagnosis method, three methods has been adopt- ed for molecular etiological research on patients of non - syndromic sensorineural hearing loss （NSHL） and their family. Methods： Six deafness genes of internal report relatively high mutation frequency were identified by restriction enzyme digestion, DNA mieroarray and Senqueneing. Results ： Among 19 cases of deafness, four cases of GJB2 235delC, one case of mt 125 rRNA A1555G were found using restriction enzyme digestion; eight patients and one of them patient＇s parents DNA microarray shows： one case of mt 12S rRNA A1555G, there cases of GJB2 235de1C, three cases of SLC26A4 gene mutations, 3 cases of 9 hot - spot mutations were normal, mt 12S rRNA A1555G and GJB2 235delc DNA microarray results are consistent with the restriction enzyme digestion. 10 cases of DNA mi- croarray detection and GJB3 sequencing were not detected GJB3 538C 〉 T mutation and other disease-causing mutation; GJB6 gene se- quences not found disease-causing mutation, mt 12S rRNA A1555G and GJB2 235delC locus sequence analysis and gene chips and di- gested again consistent with the results. SLC26A4 IVS7-2A 〉 G and 2168A 〉 G sequence analysis was consistent with the DNA mi- croarray. Followed by parents, some patients using the above method for screening, 48 cases of samples, 15 cases were detected Muta- tion-indticed deafness carrier, mutation detection rate was 31.25%. Conclusion： Three results of Enzyme digestion, DNA mieroarray corresponds to the sequencing. Three kinds of detection techniques were Integrated in accordance of patient in order to provide a molecular basis for clinical diagnosis.