129例辅助生殖技术子代儿童耳聋基因突变筛查

目的:了解辅助生殖技术(assisted reproductive technology,ART)子代儿童常见遗传性耳聋基因突变发生率。方法:随机选取2010年1月-2014年12月在中国科学技术大学附属第一医院行ART助孕成功的129例子代为研究对象,平均年龄为(4.37±0.92)岁,同时收集161例自然妊娠出生的子代血样。利用高通量测序法对研究对象的常见耳聋基因突变位点进行筛查。结果:129例ART子代共发现11例携带耳聋基因突变,包含12个基因位点的突变,而161例自然妊娠子代共发现9例携带耳聋基因突变,包含9个基因位点的突变,2组耳聋基因突变发生率比较差异无统计学意义(χ^2=0.962,P=0.327)。ART子代中GJB2突变携带者2例(1.55%),男女比例为1∶1;GJB3突变携带者3例(2.33%),男女比例为1∶2;SLC26A4突变携带者7例(5.43%),男女比例为5∶2;未发现线粒体12S r RNA基因突变携带者。1例有GJB2基因235delC位点和SLC26A4基因IVS7-2A>G位点双杂合突变。ART子代中,体外受精(IVF)组耳聋基因位点突变发生率为11.83%(11/93),胞浆内单精子注射(ICSI)组为2.78%(1/36),差异无统计学意义(χ^2=2.520,P=0.112);新鲜胚胎移植组为11.39%(9/79),冷冻胚胎移植组为6.00%(3/50),差异无统计学意义(χ^2=1.055,P=0.304)。结论:ART子代遗传性耳聋基因突变发生率与自然妊娠组子代相似;ART组以SLC26A4基因IVS7-2A>G位点突变阳性率最高。而受精方式以及移植新鲜或冻融胚胎对ART子代的遗传性耳聋的基因突变发生率无显著影响。

Screening of Common Deafness Gene Mutations in 129 Children Conceived through Assisted Reproductive Technology

Objective: To investigate the mutative rate of common hereditary deafness genes in children conceived through assisted reproductive technology(ART offspring). Methods:A total of 129 cases of ART offspring in our center from January 2010 to December 2014 were included, and the average age of the subjects was(4.37±0.92) years old. Blood samples of 161 offspring conceived through natural pregnancy(as the control group) were collected. High-throughput sequencing was used to screen the deafness gene mutation sites. Results:A total of 11 ART offspring were found to have deafness gene mutations, including 12 mutations in the gene locus.A total of 9 cases in the control group were found to carry deafness gene mutations, including 9 mutations in the gene locus. There was no statistically significance of the mutation incidence in the two groups( χ^2=0.962, P =0.327). In ART offspring, there were 2 cases of GJB2 mutation carrier(1.55%, male to female was 1∶1), 3 cases of GJB3 mutation carrier(2.33%, male to female was 1∶2), 7 cases of SLC26 A4 mutation carrier(5.43%, male to female was 5 ∶2) and one case of GJB2 gene 235 delC site and a SLC26 A4 gene IVS7-2 A >G site double heterozygous mutation. There was no mitochondrial 12 S rRNA gene mutation carrier in those ART offspring. The mutation rate of gene locus was 11.83%(11/93) in the IVF group, and 2.78%(1/36) in the ICSI group, with no significant difference between the IVF group and the ICSI group( χ^2=2.520, P=0.112). The mutation rate of gene locus was 11.39%(9/79) in the fresh embryo transfer group, and 6.00%(3/50) in the frozen embryo transfer group,with no significant difference between the two groups( χ^2=1.055, P =0.304). Conclusions:The incidence of hereditary deafness gene mutations in ART offspring is similar with the offspring conceived through natural pregnancy. In ART offspring, the positive rate of SLC26 A4 gene IVS7-2 A>G site mutation is the highest. There is no significant effect of the fertilization method and the fresh or frozen-thawed embryo transfer on the mutation rate of hereditary deafness in ART offspring.