title: Identification of novel mutations of PKD1 gene in Chinese patients with autosomal dominant polycystic kidney disease by targeted next-generation sequencing.
abstract: BACKGROUND
  Mutations of PKD1 and PKD2 accounted for the most cases of autosomal dominant polycystic kidney disease (ADPKD). The presence of the large transcript, numerous exons and complex reiterated regions within the gene has significantly complicated the analysis of PKD1 with routine PCR-based approaches.
  
  
  METHODS
  We developed a strategy to analyze both the PKD1/PKD2 genes simultaneously using targeted next-generation sequencing (NGS). All coding exons plus the flanking sequences of PKD1 and PKD2 genes from probands were captured, individually barcoded and followed by HiSeq2000 sequencing. The candidate variants were validated by using classic Sanger sequencing. PKD1-specific primers were designed to amplify the replicated areas of PKD1 gene.
  
  
  RESULTS
  Five novel variations and one known mutation in PKD1 gene were detected in five familial and one sporadic Chinese ADPKD patients. Through pedigree and bioinformatic analysis, five of them were identified as pathogenic mutations (p.G1319R, p.Y3781*, p.W4122*, p.Val700Glyfs*14, and p.Leu3656Trpfs*28) and one was as polymorphism (p.T2420I).
  
  
  CONCLUSIONS
  Our result showed that targeted capture and NGS technology were effective for the gene testing of ADPKD disorder. Mutation study of PKD1 and PKD2 genes in Chinese patients may contribute to better understanding of the genetic diversity between different ethnic groups and enrich the mutation database in Asian population.
authors.affiliation: Department of Medical Genetics, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, WHO Collaborating Centre for Community Control of Hereditary Diseases, Beijing, China. Electronic address: tyang@ibms.pumc.edu.cn.
authors.forename: Tao
authors.initials: T
authors.lastname: Yang
authors.affiliation: Department of Medical Genetics, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, WHO Collaborating Centre for Community Control of Hereditary Diseases, Beijing, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Meng
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Wei
authors.affiliation: The State Key Laboratory of Reproductive Medicine, Clinical Center of Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, China.
authors.forename: Jiandong
authors.initials: J
authors.lastname: Shen
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Mingrong
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Medical Genetics, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, WHO Collaborating Centre for Community Control of Hereditary Diseases, Beijing, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Qi
authors.affiliation: Department of Medical Genetics, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, WHO Collaborating Centre for Community Control of Hereditary Diseases, Beijing, China.
authors.forename: Chundan
authors.initials: C
authors.lastname: Wang
authors.affiliation: The State Key Laboratory of Reproductive Medicine, Clinical Center of Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, China.
authors.forename: Jiayin
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Medical Genetics, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, WHO Collaborating Centre for Community Control of Hereditary Diseases, Beijing, China.
authors.forename: Minrui
authors.initials: M
authors.lastname: Ma
authors.affiliation: Department of Medical Genetics, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, WHO Collaborating Centre for Community Control of Hereditary Diseases, Beijing, China. Electronic address: hsz_pumc@ibms.pumc.edu.cn.
authors.forename: Shangzhi
authors.initials: S
authors.lastname: Huang
mesh_terms: D000328:Adult; D000818:Animals; D044466:Asian People / Q000235:genetics*; D001483:Base Sequence; D019295:Computational Biology; D004252:DNA Mutational Analysis; D005260:Female; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D016891:Polycystic Kidney, Autosomal Dominant / Q000175:diagnosis / Q000235:genetics*; D000091283:Protein Kinase D2; D011494:Protein Kinases / Q000235:genetics; D017434:Protein Structure, Tertiary; D050396:TRPP Cation Channels / Q000737:chemistry / Q000235:genetics*; D055815:Young Adult
chemical_list: D000091283:Protein Kinase D2; D011494:Protein Kinases; D050396:TRPP Cation Channels
keywords: ADPKD; Mutation; PKD1; Targeted next-generation sequencing