Differential High-Resolution Melting Analysis for the Detection of K-ras Codons 12 and 13 Mutations in Pancreatic Cancer

作者：Zhang, Chunyan; Guo, Wei^*; Wu, Jiong; Song, Binbin; Zhang, Chen; Dai, Qian; Pan, Baishen; Ji, Yuan; Guo, Jinghui

来源：Pancreas, 2011, 40(8): 1283-1288.

DOI：10.1097/MPA.0b013e318220af91

• high-resolution melting analysis
• K-ras genes
• mutation
• pancreatic cancer

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.

• 出版日期2011-11
• 单位复旦大学