多重连接探针扩增技术诊断鸟氨酸氨甲酰转移酶缺乏症五例  

Applying multiplex ligation-dependent probe amplification in the diagnosis of 5 cases with ornithine transcarbamylase deficiency

作　　者：龚珠文[1] 韩连书[1] 叶军[1] 邱文娟[1] 张惠文[1] 余永国[1] 梁黎黎[1] 高晓岚[1] 王瑜[1] 季文君[1] 顾学范[1] 

机构地区：[1]上海交通大学医学院附属新华医院 /上海市儿科医学研究所内分泌/遗传科,200092

出　　处：《中华儿科杂志》2016年第6期437-440,共4页Chinese Journal of Pediatrics

基　　金：国家科技支撑计划（2012BA109804）

摘　　要：目的 运用多重连接探针扩增技术（MLPA）检测鸟氨酸氨甲酰转移酶（OTC）基因大片段缺失和重复突变.方法 2012年7月至2014年10月上海交通大学医学院附属新华医院串联质谱检测疑似OTC缺乏症（OTCD）的患儿30例,其中男13例、女17例.进行Sanger测序,23例明确突变.未检测到突变的疑似OTCD患儿行MLPA检测,进行DNA变性、探针与样本DNA的杂交、杂交探针的连接、连接探针的PCR扩增、PCR产物的毛细管电泳.使用Coffalyser软件分析数据.结果 5例患儿Sanger测序未检测到突变,MLPA检测发现异常.例1女,9岁,外显子2～4杂合缺失;例2男,出生后10 d死亡,抽母血检测为外显子2～6杂合重复;例3男,出生10d,外显子7～10半合子缺失;例4女,2岁,外显子1～4杂合缺失;例5男,6岁时死亡,抽母血检测为外显子1～4杂合重复.结论 MLPA检测有助于提高OTC基因致病突变的检出,对于疑似OTCD患者,Sanger测序未检测到突变的患者需要进行MLPA检测.Objective To detect large genomic deletions or duplications of ornithine transcarbamylase （OTC） gene by multiplex ligation-dependent probe amplification （MLPA）.Method Thirty cases of suspected OTC deficiency （OTCD） patients based on tandem-mass spectrum results were recruited in Xinhua Hospital from 2012 to 2014,among whom 13 were male and 17 were female.Sanger sequencing of OTC gene revealed mutations in 23 cases.MLPA was performed in the patients whose previous Sanger sequencing failed to detect any disease-causing mutation.The samples were treated via the steps of DNA degeneration,the probe hybridization,connecting the hybridization probe,PCR amplification and capillary electrophoresis.The data were analyzed using Coffalyser software.Result Abnormal MLPA results were found in 5 patients without mutation detected in previous Sanger sequencing.Patient 1,a 9-year old girl,had a heterozygous deletion of Exon 2-4.Patient 2,a male newborn,died 10 days after birth.The examination of the mother＆#39;s sample by MLPA revealed a heterozygous duplication of exon 2-6.Patient 3,a 10-day old boy,was found to harbor a hemizygous deletion of exon 7-10.Patient 4,a 2-year old girl,harbored a heterozygous deletion of exon 1-4.The fifth patient died at the age of 6 years,and his mother carried a heterozygous duplication of exon 1-4.Conclusion MLPA can be helpful in detecting the OTC gene defects,particularly for OTCD patients without mutation detected by Sanger sequencing.

关 键 词：鸟氨酸氨甲酰转移酶缺乏症 基因测定 多重连接探针扩增技术 

分 类 号：R725.9[医药卫生—儿科] R440[医药卫生—临床医学]