Suzuki Shun-Ichi, Matsusaka Satoshi, Hirai Mitsuharu, Shibata Harumi, Takagi Koichi, Mizunuma Nobuyuki, Hatake Kiyohiko
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.
Int J Oncol. 2015 Jul;47(1):97-105. doi: 10.3892/ijo.2015.2978. Epub 2015 Apr 30.
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.
据报道，患有KRAS和BRAF突变（位于表皮生长因子受体（EGFR）下游）的结肠癌患者对使用抗EGFR抗体西妥昔单抗和帕尼单抗的治疗产生耐药性。另一方面，一些报告称KRAS密码子13突变（p.G13D）对抗EGFR抗体的耐药性较低，因此迫切需要检测特定的KRAS突变。我们建立了一种使用QProbe（QP）方法的先进测量系统，该系统可以同时测量KRAS密码子12/13、p.G13D和BRAF突变，并将该方法与使用182例结肠癌患者样本的直接测序（DS）方法进行比较。此外，为了检测KRAS/BRAF，对32份活检样本采用了一种无需DNA纯化的新型预处理方法进行处理。KRAS突变测量结果显示，除5份无法确定的样本外，QP方法与DS方法之间的一致性率为81.4%（144/177）。其中，29份样本QP方法检测为阳性，DS方法检测为阴性。BRAF仅用QP方法测量，突变检出率为3.9%（6/153）。使用一种无需DNA提取的简单新预处理方法进行KRAS测量，32份样本中有31份结果良好，所有结果均与DS方法匹配。我们建立了一种简单但高度灵敏的KRAS/BRAF同时检测系统。此外，新型预处理技术的引入消除了不便的DNA提取过程。从这一研究成果来看，我们不仅期望在临床领域能得到快速准确的结果，也期望对提高检测质量和工作效率有所贡献。
