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中国循证儿科杂志

中国循证儿科杂志 ›› 2016, Vol. 11 ›› Issue (2): 131-136.

• 论著 • 上一篇    下一篇

基于快速和常规全外显子组分析技术对遗传性疾病的诊断程序比较

杨琳1,3 董辰1,3 魏泽峻2 卢宇蓝1 吴冰冰1 王慧君1 周文浩1   

  1. 1 复旦大学附属儿科医院分子诊断中心 上海，201102；2 明码（上海）生物科技有限公司 上海;3 共同第一作者
  • 收稿日期:2016-04-21 修回日期:2016-04-21 出版日期:2016-04-25 发布日期:2016-04-21
  • 通讯作者: 周文浩

Procedure comparison between rapid and standard whole-exome data interpretation for clinical diagnosis of genetic disorder

YANG Lin1,3, DONG Chen1,3, WEI Ze-jun2, LU Yu-lan1, WU Bin-bin1, WANG Hui-jun1, ZHOU Wen-hao1   

  1. 1 Children′s Hospital of Fudan University, Shanghai 201102; 2 WuXi Next CODE Genorvics(Shanghai) Co; Ltd, Shanghai 200131,China; 3 Co-first author
  • Received:2016-04-21 Revised:2016-04-21 Online:2016-04-25 Published:2016-04-21
  • Contact: ZHOU Wen-hao

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摘要：

目的 探索可被临床医生直接应用的全外显子测序（WES）数据分析平台及方法。方法 选择复旦大学附属儿科医院（我院）临床诊断不明疾病的核心家系3例（例1~3）和仅先证者2例（例4和5）行WES分析，采用WuXi NextCODE临床测序数据分析系统（简称NextCODE平台）进行快速数据分析，并与我院分子诊断中心已建立的高通量数据分析流程（简称常规数据分析流程）进行参与人员及耗时的比较。结果 例1~3基于表型相关候选基因联合遗传模式的分析方法，分别检测到FGFR2基因杂合突变、GBE1基因复合杂合突变及TBX1基因杂合突变；例4和5通过表型相关候选基因分析，分别检测到IL10RA基因纯合突变和复合杂合突变。NextCODE平台自动完成3/7个步骤,从输入表型至生成报告，WES数据分析用时30 min以内。常规数据分析流程自动完成1/7个步骤，6个人工完成步骤需要多个专业人员进行数据的筛选及解读，从输入表型至生成报告，熟练的专业人员用时2~8 h。结论 5例临床诊断不明病例通过WES明确了诊断；NextCODE是直接为临床医生所用、简单快速的WES数据分析平台，有助于协助临床医生直接利用高通量测序数据，准确锁定致病突变，提高诊断效率。

Abstract:

Objective A difficult hurdle in whole exome sequencing application is rapid data interpretation. In this study, whole-exome sequencing and WuXi NextCODE software were used to rapidly identify pathogenic mutations in 5 undiagnosed cases. Methods The exome targets of the patient′s DNA were captured with the SureSelct Human All Exon kit followed by sequencing with the Illumina HiSeq2000 platform. The WuXi NextCODE software was used for the data analysis. The detected variant was confirmed with Sanger direct sequencing. Results 5 trio families with undiagnosed probands were recruited. A heterozygous missense mutation was identified in FGFR2 gene in proband 1, compound heterozygous missense mutations in GBE1 gene in proband 2, and a heterozygous missense mutation in TBX1 gene in proband 3 by whole-exome sequencing of trio samples. A homozygous missense mutation was identified in IL10RA gene in proband 4, and compound heterozygous missense mutations in IL10RA gene in proband 5 by whole-exome sequencing of proband only. Conclusion This study clearly showed the efficacy of whole-exome sequencing and was helpful to rapid genetic diagnosis for undiagnosed cases.