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Clin Biochem. 2014 Dec;47(18):340-3. doi: 10.1016/j.clinbiochem.2014.09.013. Epub 2014 Sep 28.

High-throughput screening of extended RAS mutations based on high-resolution melting analysis for prediction of anti-EGFR treatment efficacy in colorectal carcinoma.

Author information

1
Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan; Division of Laboratory Medicine and Clinical Genetics, Chiba University Hospital, Chiba, Japan. Electronic address: ishige.t@chiba-u.jp.
2
Division of Laboratory Medicine and Clinical Genetics, Chiba University Hospital, Chiba, Japan.
3
Department of Medical Technology and Sciences, International University of Health and Welfare, Fukuoka, Japan.
4
Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan; Division of Laboratory Medicine and Clinical Genetics, Chiba University Hospital, Chiba, Japan.
5
Department of Frontier Surgery, Graduate School of Medicine, Chiba University, Chiba, Japan.
6
Department of Diagnostic Pathology, Graduate School of Medicine, Chiba University, Chiba, Japan; Department of Pathology, Chiba University Hospital, Chiba, Japan.

Abstract

OBJECTIVES:

Recent studies have demonstrated that, in advanced colorectal carcinoma (CRC) patients, extended RAS (in KRAS exons 2-4 and NRAS exons 2-4) and BRAF mutations are negative predictors for anti-EGFR treatment efficacy and negative prognostic factor, respectively. Thus, high-throughput and cost-effective methods for identification of the mutation status are required.

DESIGN AND METHODS:

We developed a PCR-high-resolution melting (HRM)-based method for screening extended RAS and BRAF mutations, and relative frequency of mutations in formalin-fixed paraffin-embedded samples of CRC was analyzed.

RESULTS:

Among 93 CRC samples, 29 harbored mutations in KRAS exon 2, and 9 harbored mutations in BRAF exon 15. Analysis of 55 KRAS exon 2 and BRAF exon 15 wild-type CRC samples identified the following mutations: 1/55 in exon 3 and 2/55 in exon 4 of KRAS; 1/55 in exon 2, 3/55 in exon 3, and 0/55 in exon 4 of NRAS.

CONCLUSIONS:

Our PCR-HRM method will enable rapid determination of the extended RAS and BRAF mutation status prior to anti-EGFR treatment in the clinical setting.

KEYWORDS:

Anti-EGFR treatment; Colorectal carcinoma; Extended RAS; High-resolution melting

[Indexed for MEDLINE]

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