应用高通量测序技术对远端肾小管酸中毒家系进行基因诊断

Genetic diagnosis of pedigrees with distal renal tubular acidosis using high throughput sequencing technology

目的对2例远端肾小管酸中毒家系进行基因突变分析。方法收集远端肾小管酸中毒患儿病历资料,应用高通量测序技术对患儿基因组DNA进行突变筛查,Sanger测序方法对患儿及家系成员进行验证,对可疑突变进行生物信息学预测。结果测序结果显示病例1患儿携带ATP6V0A4基因c.1185delC和c.1418C>T复合杂合突变;病例2患儿携带ATP6V0A4基因c.639+1G>A和c.2227C>T复合杂合突变; 2例患儿的突变均分别遗传自父母。c.1418C>T和c.2227C>T突变均未见报道,经生物信息学预测为有害突变,根据美国医学遗传学与基因组学会遗传变异分类指南均归类为可能致病性变异。结论发现ATP6V0A4基因2个新的备选突变位点,为基因检测和诊断治疗提供借鉴。

Objective To analyze the gene mutations of two pedigrees with distal renal tubular acidosis. Methods The data of children with distal renal tubular acidosis were collected,and their mutations of genomic DNA were screened by the high throughput sequencing technology. The suspected mutations were verified by Sanger sequencing,and then bioinformatics prediction was performed. Results The sequencing results showed that a heterozygous c.1185delC and c.1418C>T mutation of ATP6V0A4 gene existed in one pediatric patient and that a heterozygous c.639+1G>A and c.2227C>T mutation of ATP6V0A4 gene existed in the other. The mutations of the two pediatric patients were inherited from their parents,respectively. The mutations of c.1418C>T and c.2227C>T had not been reported yet,and they were predicted to be harmful by the bioinformatics analysis. According to the standards and guidelines of the American College of Medical Genetics (ACMG) ,the two mutations were classified as likely pathogenic variation. Conclusion Two new candidate mutation sites of ATP6V0A4 gene are found,which may be used for gene detection and genetic diagnosis and treatment.