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Pancreas. 2011 Nov;40(8):1283-8. doi: 10.1097/MPA.0b013e318220af91.

Differential high-resolution melting analysis for the detection of K-ras codons 12 and 13 mutations in pancreatic cancer.

Author information

1
Department of Laboratory Medicine, Zhongshan Hospital, Fudan University, Shanghai, China.

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.

PMID:
21989023
DOI:
10.1097/MPA.0b013e318220af91
[Indexed for MEDLINE]

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