title: High-resolution melting analyses for gene scanning of APC, MLH1, MSH2, and MSH6 associated with hereditary colorectal cancer.
abstract: BACKGROUND
  Hereditary colorectal cancer accounts for approximately 4-5% of all colorectal cancers. The causative genes for familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer are large, making comprehensive analyses difficult. Therefore, high-throughput and practical methods are required to make an early diagnosis of hereditary colorectal cancers and identify high-risk individuals. For this purpose, we developed a novel gene scanning method by high-resolution melting (HRM) analysis.
  
  
  METHODS
  High-resolution melting (HRM) analysis is a promising prescreening method for nucleic acid sequence variants because of its high sensitivity and high-throughput capability. We evaluated HRM for screening APC, MLH1, MSH2, and MSH6 genes for point mutations, small deletions, and insertions. Simultaneously, we evaluated quantitative polymerase chain reaction-HRM (qPCR-HRM) for screening the MSH2 gene for large rearrangements.
  
  
  RESULTS
  All 28 point mutations and 1 large rearrangement were successfully detected by qPCR-HRM analysis.
  
  
  CONCLUSIONS
  A fast and reliable mutation detection strategy with HRM and qPCR-HRM was used to diagnose hereditary colorectal cancers. Because this method is simple and economical, it may be useful in diagnostic laboratories.
authors.affiliation: Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan. jurat0922@yahoo.co.jp
authors.forename: Jurat
authors.initials: J
authors.lastname: Obul
authors.forename: Sakae
authors.initials: S
authors.lastname: Itoga
authors.forename: Maynur
authors.initials: M
authors.lastname: Abliz
authors.forename: Kenichi
authors.initials: K
authors.lastname: Sato
authors.forename: Takayuki
authors.initials: T
authors.lastname: Ishige
authors.forename: Emi
authors.initials: E
authors.lastname: Utsuno
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Matsushita
authors.forename: Hisahiro
authors.initials: H
authors.lastname: Matsubara
authors.forename: Fumio
authors.initials: F
authors.lastname: Nomura
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D025601:Adenomatous Polyposis Coli Protein / Q000235:genetics; D003123:Colorectal Neoplasms, Hereditary Nonpolyposis / Q000175:diagnosis* / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics; D015321:Gene Rearrangement; D006801:Humans; D000070957:MutL Protein Homolog 1; D051718:MutS Homolog 2 Protein / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics; D009691:Nucleic Acid Denaturation; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D044366:Transition Temperature
chemical_list: C555337:APC protein, human; D048868:Adaptor Proteins, Signal Transducing; D025601:Adenomatous Polyposis Coli Protein; D004268:DNA-Binding Proteins; C094443:G-T mismatch-binding protein; C501486:MLH1 protein, human; D009687:Nuclear Proteins; C497172:MSH2 protein, human; D000070957:MutL Protein Homolog 1; D051718:MutS Homolog 2 Protein