title: Rapid Detection Method for the Four Most Common CHEK2 Mutations Based on Melting Profile Analysis.
abstract: BACKGROUND
  CHEK2 is a tumor suppressor gene, and the mutations affecting the functionality of the protein product increase cancer risk in various organs. The elevated risk, in a significant percentage of cases, is determined by the occurrence of one of the four most common mutations in the CHEK2 gene, including c.470T>C (p.I157T), c.444+1G>A (IVS2+1G>A), c.1100delC, and c.1037+1538_1224+328del5395 (del5395).
  
  
  METHODS
  We have developed and validated a rapid and effective method for their detection based on high-resolution melting analysis and comparative-high-resolution melting, a novel approach enabling simultaneous detection of copy number variations. The analysis is performed in two polymerase chain reactions followed by melting analysis, without any additional reagents or handling other than that used in standard high-resolution melting.
  
  
  RESULTS
  Validation of the method was conducted in a group of 103 patients with diagnosed breast cancer, a group of 240 unrelated patients with familial history of cancer associated with the CHEK2 gene mutations, and a 100-person control group. The results of the analyses for all three groups were fully consistent with the results from other methods.
  
  
  CONCLUSIONS
  The method we have developed improves the identification of the CHEK2 mutation carriers, reduces the cost of such analyses, as well as facilitates their implementation. Along with the increased efficiency, the method maintains accuracy and reliability comparable to other more labor-consuming techniques.
authors.affiliation: Institute of Human Genetics, Polish Academy of Sciences, Ul. Strzeszyńska 32, 60-479, Poznan, Poland.
authors.forename: Pawel
authors.initials: P
authors.lastname: Borun
authors.affiliation: Institute of Human Genetics, Polish Academy of Sciences, Ul. Strzeszyńska 32, 60-479, Poznan, Poland.
authors.forename: Kacper
authors.initials: K
authors.lastname: Salanowski
authors.affiliation: Cancer Prevention and Epidemiology Center Poznan, Poznan, Poland.
authors.forename: Dariusz
authors.initials: D
authors.lastname: Godlewski
authors.affiliation: Department of Pediatric Gastroenterology and Metabolic Diseases, University of Medical Sciences, Poznan, Poland.
authors.forename: Jaroslaw
authors.initials: J
authors.lastname: Walkowiak
authors.affiliation: Institute of Human Genetics, Polish Academy of Sciences, Ul. Strzeszyńska 32, 60-479, Poznan, Poland. andp@man.poznan.pl.|Genetic Diagnostics Center GENESCREEN, Kobylnica, Poland. andp@man.poznan.pl.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Plawski
mesh_terms: D001943:Breast Neoplasms / Q000175:diagnosis* / Q000235:genetics; D016022:Case-Control Studies; D064447:Checkpoint Kinase 2 / Q000235:genetics*; D056915:DNA Copy Number Variations; D004252:DNA Mutational Analysis; D005260:Female; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D025202:Molecular Diagnostic Techniques; D017384:Sequence Deletion; D044366:Transition Temperature
chemical_list: D064447:Checkpoint Kinase 2; C578634:CHEK2 protein, human