title: [Utilization of high-resolution melting analysis to screen patients with neonatal intrahepatic cholestasis caused by citrin deficiency].
abstract: OBJECTIVE
  To assess the feasibility of high-resolution melting (HRM) analysis for screening patients with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD).
  
  
  METHODS
  Based on previous studies on SLC25A13 gene in Chinese patients with NICCD, four hotspot mutations (851del4, 1638ins23, IVS6+5G>A and IVS16ins3kb) were selected. Results of the HRM analysis was validated using 50 negative controls and 20 patients with NICCD whose genotypes were confirmed previously by direct sequencing. With the established protocol, 171 suspected patients were enrolled. Samples with abnormal melting curves were further validated by DNA sequencing.
  
  
  RESULTS
  HRM analysis can accurately determine the genotypes of all negative controls and patients. The sensitivity and specificity of the technique reached 100% (70/70). The melting curves of samples with the same genotype were highly reproducible. In 171 suspected patients, seven NICCD patients were detected by HRM. Identified mutations have included one case of 851del4 homozygote, one case of IVS6+5G>A heterozygote, 3 cases of 851del4 heterozygotes, one case of [IVS6+5G>A]+[ 851del4] and one case of [1638ins23+IVS16ins3kb]+[1638ins23]. All mutations were subsequently confirmed by DNA sequencing.
  
  
  CONCLUSIONS
  HRM analysis is a convenient, high-throughput and rapid technique for the screening of NICCD patients.
authors.affiliation: Shenzhen Institute of Pediatrics, Shenzhen Children's Hospital, Shenzhen, Guangdong, People's Republic of China.
authors.forename: Peng-qiang
authors.initials: PQ
authors.lastname: Wen
authors.forename: Guo-bing
authors.initials: GB
authors.lastname: Wang
authors.forename: Zhan-ling
authors.initials: ZL
authors.lastname: Chen
authors.forename: Dong
authors.initials: D
authors.lastname: Cui
authors.forename: Xiao-hong
authors.initials: XH
authors.lastname: Liu
authors.forename: Li-fang
authors.initials: LF
authors.lastname: Ying
authors.forename: Ping
authors.initials: P
authors.lastname: Song
authors.forename: Quan
authors.initials: Q
authors.lastname: Yuan
authors.forename: Shu-li
authors.initials: SL
authors.lastname: Chen
authors.forename: Jian-xiang
authors.initials: JX
authors.lastname: Liao
mesh_terms: D027321:Anion Transport Proteins / Q000235:genetics; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000172:deficiency*; D002681:China; D020159:Citrullinemia / Q000175:diagnosis* / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D020022:Genetic Predisposition to Disease; D005838:Genotype; D006801:Humans; D024101:Mitochondrial Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D009691:Nucleic Acid Denaturation; D027361:Organic Anion Transporters / Q000172:deficiency*; D012680:Sensitivity and Specificity
chemical_list: D027321:Anion Transport Proteins; D002135:Calcium-Binding Proteins; D024101:Mitochondrial Proteins; D027361:Organic Anion Transporters; C532592:Slc25a1 protein, human; C008822:citrin; D004247:DNA