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首页> 美国卫生研究院文献>International Journal of Oncology >A novel approach to detect KRAS/BRAF mutation for colon cancer: Highly sensitive simultaneous detection of mutations and simple pre-treatment without DNA extraction
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A novel approach to detect KRAS/BRAF mutation for colon cancer: Highly sensitive simultaneous detection of mutations and simple pre-treatment without DNA extraction

机译：一种检测结肠癌KRAS / BRAF突变的新方法：高度敏感的同时检测突变和无需DNA提取的简单预处理

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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-theart 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.
机译：据报道，位于表皮生长因子受体（EGFR）下游的具有KRAS和BRAF突变的结肠癌患者获得抗EGFR抗体，西妥昔单抗和帕尼单抗治疗的耐药性。另一方面，一些报道说KRAS密码子13突变（p.G13D）对抗EGFR抗体的抵抗力较低，因此非常需要检测特定的KRAS突变。我们已经建立了使用QProbe（QP）方法的最新测量系统，该系统可以同时测量KRAS密码子12/13，p.G13D和BRAF突变，并且使用来自结肠的182个标本将该方法与直接测序（DS）进行了比较癌症患者。另外，用新的预处理方法对32个活检标本进行了处理，而无需进行DNA纯化，以检测KRAS / BRAF。 KRAS突变测量的结果是，QP法和DS法之间的一致性率为81.4％（144/177），除了5个标本不确定。其中，有29个样本的QP方法呈阳性，DS方法呈阴性。仅使用QP方法测量BRAF，突变检测率为3.9％（6/153）。使用简单的新型预处理方法进行的KRAS测量无需DNA提取即可获得32个结果中的31个良好结果，所有这些结果均与DS方法匹配。我们已经为KRAS / BRAF建立了一个简单但高度灵敏的同时检测系统。此外，新型预处理技术的引入消除了不方便的DNA提取过程。从这项研究成果中，我们不仅可以期望在临床领域获得快速，准确的结果，而且可以为提高测试质量和工作效率做出贡献。

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