title: [Large deletion in mismatch repair genes uncovered by quantitative multiplex PCR-high performance liquid chromatography system].
abstract: OBJECTIVE
  Establishing a new method on the basis of multiplex PCR-high performance liquid chromatography (HPLC) for screening a large deletion in mismatch repair genes.
  
  
  METHODS
  Thirty-five pairs of primers were used to amplify all 16 exons of MSH2 and all 19 exons of MLH1 gene in 8 multiplex PCR. The products of multiplex PCR were analysed for the large deletion with Double Strand DNA Analysis System of HPLC. Firstly, validation of the method was tested on positive and negative controls in blind analysis. Secondly, 14 blood cell DNA samples from hereditary nonpolyposis colorectal cancer (HNPCC) patients and 13 colorectal cancer (CRC) tissues DNA samples from sporadic CRC patients were checked with the new developed method.
  
  
  RESULTS
  (1) the genomic deletions in all 4 of positive controls were identically uncovered with the new method; (2) a novel germline and a novel somatic large deletions were unveiled in 1/14 HNPCC patients and in 1/13 CRC tissues.
  
  
  CONCLUSIONS
  The method developed on multiplex PCR-HPLC is reliable for uncovering large genomic deletion in mismatch repair genes, and can be taken as a valuable addition to mutation screening system.
authors.affiliation: Department of Molecular Biology, Jiangsu Institute of Cancer Research, Nanjing, Jiangsu, 210009 PR China. wangyapn@jlonline.com
authors.forename: Ya-ping
authors.initials: YP
authors.lastname: Wang
authors.forename: Ming
authors.initials: M
authors.lastname: Zhu
authors.forename: Meng-hong
authors.initials: MH
authors.lastname: Sun
authors.forename: Jin-tian
authors.initials: JT
authors.lastname: Li
authors.forename: Jun-ni
authors.initials: JN
authors.lastname: Zhang
authors.forename: Xiao-mei
authors.initials: XM
authors.lastname: Zhang
authors.forename: Xiao-rong
authors.initials: XR
authors.lastname: Liu
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D020137:Base Pair Mismatch / Q000235:genetics*; D001483:Base Sequence; D002352:Carrier Proteins; D002851:Chromatography, High Pressure Liquid; D004260:DNA Repair / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D017353:Gene Deletion; D006801:Humans; D008969:Molecular Sequence Data; D000070957:MutL Protein Homolog 1; D051718:MutS Homolog 2 Protein; D009363:Neoplasm Proteins / Q000235:genetics*; D009687:Nuclear Proteins; D016133:Polymerase Chain Reaction / Q000379:methods*; D011518:Proto-Oncogene Proteins / Q000235:genetics*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; C501486:MLH1 protein, human; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; C497172:MSH2 protein, human; D000070957:MutL Protein Homolog 1; D051718:MutS Homolog 2 Protein