title: Multiplex tetra-primer amplification refractory mutation system PCR to detect 6 common germline mutations of the MUTYH gene associated with polyposis and colorectal cancer.
abstract: BACKGROUND
  We describe a simple tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR) for detecting MUTYH mutations, which are associated with colorectal adenomas and colorectal cancer.
  
  
  METHODS
  We designed specific T-ARMS-PCR assays for 6 mutations (Y165C, G382D, 1395_7delGGA, Y90X, 1103delC, and R231H) selected on the basis of the frequency of their occurrence. We also designed a set of 3 multiplex T-ARMS PCR assays, each for detection of 2 mutations. We tested DNA samples from patients with attenuated or classic adenomatous polyposis coli and no detectable APC germline mutations.
  
  
  RESULTS
  All mutations were easily detected with both the specific and multiplex T-ARMS-PCR assays. Results were confirmed by DNA HPLC analysis in all 54 patients, and each mutation was confirmed by direct DNA sequencing.
  
  
  CONCLUSIONS
  T-ARMS-PCR does not require any special equipment, and it provides rapid, reproducible, and cost-effective detection of common MUTYH mutations. Multiplex T-ARMS-PCR allows the detection of 6 common MUTYH mutations with use of as few as 3 single tube PCR reactions. It could be useful to carry out large population-based epidemiologic studies.
authors.affiliation: Laboratory of Human Genetics, Medical Oncology C, Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Piccioli
authors.forename: Martina
authors.initials: M
authors.lastname: Serra
authors.forename: Viviana
authors.initials: V
authors.lastname: Gismondi
authors.forename: Simona
authors.initials: S
authors.lastname: Pedemonte
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Loiacono
authors.forename: Sonia
authors.initials: S
authors.lastname: Lastraioli
authors.forename: Lucio
authors.initials: L
authors.lastname: Bertario
authors.forename: Maria
authors.initials: M
authors.lastname: De Angioletti
authors.forename: Liliana
authors.initials: L
authors.lastname: Varesco
authors.forename: Rosario
authors.initials: R
authors.lastname: Notaro
mesh_terms: D011125:Adenomatous Polyposis Coli / Q000235:genetics*; D015179:Colorectal Neoplasms / Q000235:genetics*; D045647:DNA Glycosylases / Q000235:genetics*; D018095:Germ-Line Mutation; D006801:Humans; D016133:Polymerase Chain Reaction
chemical_list: D045647:DNA Glycosylases; C061738:mutY adenine glycosylase