title: Differential high-resolution melting analysis for the detection of K-ras codons 12 and 13 mutations in pancreatic cancer.
abstract: OBJECTIVE
  The aim of this study was to establish a differential high-resolution melting analysis for the detection of K-ras codons 12 and 13 mutations in pancreatic cancer tissue.
  
  
  METHODS
  We tested the sensitivity of this genotyping approach in cell lines with known K-ras mutations using 163- and 59-base pair (bp) amplicons spanning codons 12 and 13. We then screened 50 pancreatic cancer tissues, which were subsequently sequenced to confirm mutations discovered in K-ras codons 12 and 13.
  
  
  RESULTS
  High-resolution melting analysis was more sensitive in detecting mutations than standard sequencing and was able to reliably detect as little as 10% of mutant cell line DNA diluted in wild-type cell line DNA. Compared with the 163-bp amplicons, the shorter 59-bp amplicons displayed a higher sensitivity, even at 3% to 5% dilution.
  
  
  CONCLUSIONS
  High-resolution melting analysis is a sensitive and accurate screening methodology for K-ras codons 12 and 13 mutations in clinical samples. The accuracy and sensitivity of high-resolution melting analysis can provide appropriate and cost-effective therapeutic choices for physicians.
authors: Department of Laboratory Medicine, Zhongshan Hospital, Fudan University, Shanghai, China.
authors: Chunyan
authors: C
authors: Zhang
authors: Wei
authors: W
authors: Guo
authors: Jiong
authors: J
authors: Wu
authors: Binbin
authors: B
authors: Song
authors: Chen
authors: C
authors: Zhang
authors: Qian
authors: Q
authors: Dai
authors: Baishen
authors: B
authors: Pan
authors: Yuan
authors: Y
authors: Ji
authors: Jinghui
authors: J
authors: Guo
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D003062:Codon / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D004273:DNA, Neoplasm / Q000737:chemistry / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009691:Nucleic Acid Denaturation; D010190:Pancreatic Neoplasms / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D018631:ras Proteins / Q000235:genetics*
chemical_list: D003062:Codon; D004273:DNA, Neoplasm; D018631:ras Proteins