citrin缺陷所致新生儿肝内胆汁淤积症的SLC25A13基因IVS16ins3kb突变类型分析  

Analysis of SLC25A13 gene IVS16ins3kb variation in neonatal intrahepatic cholestasis caused by citrin deficiency

作　　者：王大燕[1] 李小兵[1] 赖盼建 Wang Dayan;Li Xiaobing;Lai Panjian(Department of Pediatrics,Affiliated Jinhua Hospital,Zhejiang University School of Medicine,Jinhua 321000,Zhejiang Province,China)

机构地区：[1]浙江大学医学院附属金华医院儿科,金华321000

出　　处：《中华妇幼临床医学杂志（电子版）》2022年第3期315-322,共8页Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition)

基　　金：浙江省医药卫生科技计划项目(2020KY342);金华市科技计划项目社会发展类重点项目(2019-3-010)。

摘　　要：目的探讨citrin缺陷所致新生儿肝内胆汁淤积症(NICCD)患儿SLC25A13基因IVS16ins3kb突变类型的第二代测序技术(NGS)数据分析特点。方法选择2017年7月与2020年4月,浙江大学医学院附属金华医院儿科收治的符合NICCD临床诊断标准,采用染色体组分析工具箱(GATK)、XHMM、CNVkit软件,对患儿NGS靶向外显子捕获测序数据进行单核苷酸变异(SNV)、插入与缺失突变、拷贝数变异(CNV)分析,仅提示SLC25A13基因单个位点杂合突变的2例患儿(患儿1、2)为研究对象。采用回顾性分析方法,收集患儿1、2的临床病例资料,包括病史、实验室检查结果、基因检测结果、治疗与预后等。利用福君基因动态实验室信息管理系统3.0(FLIMS系统),采用split read方法,对患儿1、2的NGS靶向外显子捕获测序原始数据(fastq格式)进行IVS16ins3kb突变类型分析,并采用长链、高保真PCR(LA-PCR)方法进行验证。本研究遵循的程序符合浙江大学医学院附属金华医院医学伦理委员会规定,并获得该委员会批准(审批文号:2018-119)。结果①病史采集:患儿1(女性)与患儿2(男性),分别因皮肤黄染2个月余与4个月余,于本院就诊,就诊时年龄分别为2个月+19 d、4个月+16 d,均为足月儿、均无家族遗传性疾病史。②实验室检查、治疗:入院时,对患儿1、2干血片采取串联质谱仪进行遗传代谢病检测结果显示,血液中瓜氨酸、甲硫氨酸等多种氨基酸含量显著增高;肘静脉血生化检查结果显示,血清氨、乳酸浓度及血清总胆红素(STB)、血清直接胆红素(SDB)、血清间接胆红素(SIB)、天冬氨酸氨基转移酶(AST)、γ-谷氨酰转移酶、碱性磷酸酶均异常增高。对患儿1、2均采取无乳糖配方奶喂养3个月后,血液生化指标均明显好转。③NGS靶向外显子捕获及Sanger测序法验证结果:采用GATK、XHMM、CNVki软件对患儿1、2的NGS靶向外显子捕获测序数据进行SNV、插入与缺失突变、CNV�Objective To explore characteristics of next-generation sequencing(NGS)data analysis of SLC25A13 gene IVS16ins3kb variation in children with neonatal intrahepatic cholestasis caused by citrin deficiency(NICCD).Methods On July 2017 and April 2020,two cases of clinically diagnosed NICCD(case 1 and case 2)who had only single locus heterozygous variation of SLC25A13 gene confirmed by single nucleotide variation(SNV),insertion and deletion variations,and copy number variation(CNV)analysis of NGS targeted exon capture sequencing data using genome analysis toolbox(GATK),XHMM,and CNVkit softwares were selected as research subjects.Clinical data of 2 children,including medical history,laboratory test results,genetic test results,treatment and prognosis were collected by retrospective analysis method.With use of Fujun Gene FLIMS system(Fujun Gene Dynamic Laboratory Information Management System 3.0),split read method was used to analyzed IVS16ins3kb variation in original data of NGS targeted exon capture sequencing(fastq format)in case 1 and case 2,and long and accurate PCR(LA-PCR)method was used to prove IVS16ins3kb variation.This study was approved by the Medical Ethics Committee of Affiliated Jinhua Hospital,Zhejiang University School of Medicine(Approval No.2018-119).Results①Medical history:case 1(female)and case 2(male)were treated in our hospital due to xanthochromia for more than 2 months and more than 4 months,respectively.Their ages at admission to hospital were 2-month-19-day and 4-month-16-day,respectively.Both two were full-term infants and had no family history of hereditary diseases.②Laboratory test results,treatment and prognosis:citrulline,methionine and other amino acids of case 1 and case 2 were significantly increased by blood stasis tandem mass spectrometry genetic metabolic disease test.Serum ammonia and lactate concentrations were increased,and serum total bilirubin(STB),serum direct bilirubin(SDB),serum indirect bilirubin(SIB),aspartate aminotransferase(AST),γ-glutamyl transpeptidase,alkaline ph

关 键 词：希特林蛋白缺乏症 胆汁淤积 肝内 高通量核苷酸序列分析 SLC25A13基因 IVS16ins3kb突变 串联质谱法 分裂读片段法 儿童 

分 类 号：R722.1[医药卫生—儿科]