论文部分内容阅读
目的分析93例耳聋患儿耳聋基因携带情况,为预防遗传性耳聋提供依据。方法对绍兴市耳聋康复中心确诊的93例耳聋患儿,采用基质辅助激光解吸电离飞行时间质谱技术(MALDI-TOF-MS)检测4个常见耳聋基因GJB2、GJB3、MT-RNR1和SLC26A4的20个热点突变;采用Sanger测序法检测突变基因的全外显子,对Sanger测序无法解决的部分则采用长片段PCR、Gap PCR和定量PCR进行检测。结果 93例耳聋患儿经MALDI-TOF-MS检测,其中48例检出耳聋基因突变,检出率为51.61%。其中GJB2基因突变35例,检出率为37.63%,分别为致病突变24例,杂合突变11例;SLC26A4基因突变13例,检出率为13.98%,分别为SLC26A4致病突变6例,杂合突变7例;未检出MT-RNR1和GJB3基因突变。对18例检出杂合突变患儿中的17例进行突变杂合基因的全外显子检测,结果检出12例(70.59%)存在其他位点的杂合突变。最终42例患儿检出耳聋致病基因,检出率为45.16%。结论 93例耳聋患儿中,GJB2、SLC26A4基因检出率较高,对常规筛查中发现的携带杂合突变基因患儿进行该基因的全外显子检测,有助于提高耳聋致病基因检出率。
Objective To analyze 93 cases of deafness in children with deafness gene carrier, to provide the basis for the prevention of hereditary deafness. Methods Ninety-three children with deafness diagnosed by Deafness Rehabilitation Center of Shaoxing City were enrolled. Twenty children with deafness gene GJB2, GJB3, MT-RNR1 and SLC26A4 were detected by matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF- Hot spot mutations were detected. Sanger sequencing was used to detect all exons of the mutated genes. Long-sequence PCR, Gap PCR and quantitative PCR were used to detect the unsolvable parts of Sanger sequencing. Results 93 children with deafness were detected by MALDI-TOF-MS, of which 48 cases were detected deafness gene mutation, the detection rate was 51.61%. GJB2 gene mutation in 35 cases, the detection rate was 37.63%, respectively, pathogenic mutation in 24 cases, 11 cases of heterozygous mutation; SLC26A4 gene mutation in 13 cases, the detection rate was 13.98%, respectively, SLC26A4 pathogenic mutations in 6 cases, 7 cases of heterozygous mutation; no mutation of MT-RNR1 and GJB3 gene was detected. Seventeen out of 18 children with heterozygous mutation were detected for all exons of the mutant heterozygous gene, and 12 (70.59%) heterozygous mutations were found in other sites. The final 42 cases of children detected deafness gene, the detection rate was 45.16%. Conclusion The detection rate of GJB2 and SLC26A4 genes in 93 children with deafness is high. Detection of all exons of this gene in children with heterozygous mutation found in routine screening can improve the gene expression of deafness The detection rate.