title: [A PKLR Gene Novel Complex Mutation in Erythrocyte Pyruvate Kinase Deficiency Detected by Targeted Sequence Capture and Next Generation Sequencing].
abstract: OBJECTIVE
  To explore the molecular mechanism of erythrocyte pyruvate kinase deficiency (PKD).
  
  
  METHODS
  Targeted sequence capture and next-generation sequencing (NGS) were used to detect the regions of exon and exon-intron boundarie of PKLR gene in a clinical suspected PKD patient. The protein function of mutant gene was forecasted by the SIFT and PolyPhen-2 databank, after the mutation of PKLR gene in the patient was detected by the NGS technology, its genotype was confirmed by Sanger sequencing.
  
  
  RESULTS
  The patient was found to have peculiar double heterozygous mutations: 661 G>A (Asp221Asn) of exon 5 and 1528 C>T (Arg510Ter) of exon 10, resulting in amino acid substitution Asp221Asn and Arg510Ter, these mutations were also further confirmed by Sanger sequencing. The complex mutations were infrequent and each of them was able to cause diseases.
  
  
  CONCLUSIONS
  The complex mutations of both 661 G>A and 1528 C>T of PKLR gene are the molecular mechanism of PKD. Simultaneous existance of above-mentioned complex mutations in PDK patient was never been previously reported at home and abroad.
authors.affiliation: Department of Hematology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang 050082, Hebei Province, China. E-mail: ldle2008@sina.com.
authors.forename: Dong-Liang
authors.initials: DL
authors.lastname: Li
authors.affiliation: Department of Internal Medicine, Shijiazhuang Hospital of Integrated Traditional Chinese and Western Medicine, Shijiazhuang 050000, Hebei Province, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Hematology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang 050082, Hebei Province, China.
authors.forename: Yan-Li
authors.initials: YL
authors.lastname: Liu
authors.affiliation: Department of Hematology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang 050082, Hebei Province, China.
authors.forename: Bao-Quan
authors.initials: BQ
authors.lastname: Jiao
authors.affiliation: Department of Hematology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang 050082, Hebei Province, China.
authors.forename: Zhi-Wei
authors.initials: ZW
authors.lastname: Wang
authors.affiliation: Department of Hematology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang 050082, Hebei Province, China.
authors.forename: You-Jun
authors.initials: YJ
authors.lastname: Wang
authors.affiliation: Department of Hematology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang 050082, Hebei Province, China.
authors.forename: Wen-Jing
authors.initials: WJ
authors.lastname: Li
authors.affiliation: Department of Hematology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang 050082, Hebei Province, China.
authors.forename: Lan-Fen
authors.initials: LF
authors.lastname: Hou
authors.affiliation: Department of Hematology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang 050082, Hebei Province, China.
authors.forename: Hong-Mou
authors.initials: HM
authors.lastname: Guo
authors.affiliation: Department of Hematology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang 050082, Hebei Province, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Hematology, Bethune International Peace Hospital of Chinese PLA, Shijiazhuang 050082, Hebei Province, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Guo
mesh_terms: D000746:Anemia, Hemolytic, Congenital Nonspherocytic / Q000235:genetics*; D005091:Exons; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007438:Introns; D009154:Mutation; D011770:Pyruvate Kinase / Q000172:deficiency* / Q000235:genetics*; D015323:Pyruvate Metabolism, Inborn Errors / Q000235:genetics*
chemical_list: D011770:Pyruvate Kinase