A novel approach to detect KRAS/BRAF mutation for colon cancer: Highly sensitive simultaneous detection of mutations and simple pre-treatment without DNA extraction

Suzuki,Shun-Ichi; Matsusaka,Satoshi; Hirai,Mitsuharu; Shibata,Harumi; Takagi,Koichi; Mizunuma,Nobuyuki; Hatake,Kiyohiko

doi:10.3892/ijo.2015.2978

A novel approach to detect KRAS/BRAF mutation for colon cancer: Highly sensitive simultaneous detection of mutations and simple pre-treatment without DNA extraction

Authors:
- Shun-Ichi Suzuki
- Satoshi Matsusaka
- Mitsuharu Hirai
- Harumi Shibata
- Koichi Takagi
- Nobuyuki Mizunuma
- Kiyohiko Hatake
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Affiliations: ARKRAY Marketing Inc., Marketing Division, Shijuku-ku, Tokyo 160-0004, Japan, Gastroenterological Center, Cancer Institute Hospital of Japanese Foundation for Cancer Research, Koto-ku, Tokyo 135-8550, Japan, ARKRAY Inc., Research and Development Division, Kamigyou-ku, Kyoto 602-0008, Japan, Cancer Chemotherapy Center, Clinical Chemotherapy of Japanese Foundation for Cancer Research, Koto-ku, Tokyo 135-8550, Japan
Published online on: April 30, 2015 https://doi.org/10.3892/ijo.2015.2978
Pages: 97-105
Copyright: © Suzuki et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

It has been reported that colon cancer patients with KRAS and BRAF mutations that lie downstream of epidermal growth factor receptor (EGFR) acquire resistance against therapy with anti‑EGFR antibodies, cetuximab and panitumumab. On the other hand, some reports say KRAS codon 13 mutation (p.G13D) has lower resistance against anti-EGFR antibodies, thus there is a substantial need for detection of specific KRAS mutations. We have established a state-of-the-art measurement system using QProbe (QP) method that allows simultaneous measurement of KRAS codon 12/13, p.G13D and BRAF mutation, and compared this method against Direct Sequencing (DS) using 182 specimens from colon cancer patients. In addition, 32 biopsy specimens were processed with a novel pre-treatment method without DNA purification in order to detect KRAS/BRAF. As a result of KRAS mutation measurement, concordance rate between the QP method and DS method was 81.4% (144/177) except for the 5 specimens that were undeterminable. Among them, 29 specimens became positive with QP method and negative with DS method. BRAF was measured with QP method only, and the mutation detection rate was 3.9% (6/153). KRAS measurement using a simple new pre-treatment method without DNA extraction resulted in 31 good results out of 32, all of them matching with the DS method. We have established a simple but highly sensitive simultaneous detection system for KRAS/BRAF. Moreover, introduction of the novel pre-treatment technology eliminated the inconvenient DNA extraction process. From this research achievement, we not only anticipate quick and accurate results returned in the clinical field but also contribution in improving the test quality and work efficiency.

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