title: Targeted next-generation sequencing identification of mutations in patients with disorders of sex development.
abstract: BACKGROUND
  The identification of causative mutations is important for treatment decisions and genetic counseling of patients with disorders of sex development (DSD). Here, we designed a new assay based on targeted next-generation sequencing (NGS) to diagnose these genetically heterogeneous disorders.
  
  
  METHODS
  All coding regions and flanking sequences of 219 genes implicated in DSD were designed to be included on a panel. A total of 45 samples were used for sex chromosome dosage validation by targeted sequencing using the NGS platform. Among these, 21 samples were processed to find the causative mutation.
  
  
  RESULTS
  The sex chromosome dosages of all 45 samples in this assay were concordant with their corresponding karyotyping results. Among the 21 DSD patients, a total of 11 mutations in SRY, NR0B1, AR, CYP17A1, GK, CHD7, and SRD5A2 were identified, including five single nucleotide variants, three InDels, one in-frame duplication, one SRY-positive 46,XX, and one gross duplication with an estimated size of more than 427,038 bp containing NR0B1 and GK. We also identified six novel mutations: c.230_231insA in SRY, c.7389delA in CHD7, c.273C>G in NR0B1, and c.2158G>A, c.1825A>G, and c.2057_2065dupTGTGTGCTG in AR.
  
  
  CONCLUSIONS
  Our assay was able to make a genetic diagnosis for eight DSD patients (38.1%), and identified variants of uncertain clinical significance in the other three cases (14.3%). Targeted NGS is therefore a comprehensive and efficient method to diagnose DSD. This work also expands the pathogenic mutation spectrum of DSD.
authors.affiliation: Department of Obstetrics & Gynecology, Southwest Hospital, Third Military Medical University, Chongqing, China.
authors.forename: Yanling
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Binhai Genomics Institute, BGI-Tianjin, Tianjin, China.|BGI-Shenzhen, Shenzhen, China.|Tianjin Enterprise Key Laboratory of Clinical Molecular Diagnostic, Tianjin, China.
authors.forename: Yuting
authors.initials: Y
authors.lastname: Yi
authors.affiliation: Department of Obstetrics & Gynecology, Southwest Hospital, Third Military Medical University, Chongqing, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Yao
authors.affiliation: Binhai Genomics Institute, BGI-Tianjin, Tianjin, China.
authors.forename: Ziying
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Department of Obstetrics & Gynecology, Southwest Hospital, Third Military Medical University, Chongqing, China.
authors.forename: Huamei
authors.initials: H
authors.lastname: Hu
authors.affiliation: Binhai Genomics Institute, BGI-Tianjin, Tianjin, China.
authors.forename: Jiucheng
authors.initials: J
authors.lastname: Liu
authors.affiliation: Binhai Genomics Institute, BGI-Tianjin, Tianjin, China.
authors.forename: Changxin
authors.initials: C
authors.lastname: Gao
authors.affiliation: Binhai Genomics Institute, BGI-Tianjin, Tianjin, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Binhai Genomics Institute, BGI-Tianjin, Tianjin, China.
authors.forename: Liying
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Binhai Genomics Institute, BGI-Tianjin, Tianjin, China.|BGI-Shenzhen, Shenzhen, China.|Tianjin Enterprise Key Laboratory of Clinical Molecular Diagnostic, Tianjin, China.
authors.lastname: Asan
authors.affiliation: BGI-Shenzhen, Shenzhen, China. yix@genomics.cn.
authors.forename: Xin
authors.initials: X
authors.lastname: Yi
authors.affiliation: Department of Obstetrics & Gynecology, Southwest Hospital, Third Military Medical University, Chongqing, China. zhi.lzliang@gmail.com.
authors.forename: Zhiqing
authors.initials: Z
authors.lastname: Liang
mesh_terms: D044466:Asian People / Q000235:genetics; D002681:China; D012734:Disorders of Sex Development / Q000175:diagnosis / Q000235:genetics*; D005260:Female; D005820:Genetic Testing; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008297:Male; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA / Q000379:methods; D012730:Sex Chromosomes / Q000235:genetics; D046468:Sexual Development / Q000235:genetics
keywords: Disorders of sex development; Novel mutation; Targeted next-generation sequencing