TY - JOUR T1 - <em>KRAS</em>, <em>NRAS</em> and <em>BRAF</em> mutations in Greek and Romanian patients with colorectal cancer: a cohort study JF - BMJ Open JO - BMJ Open DO - 10.1136/bmjopen-2013-004652 VL - 4 IS - 5 SP - e004652 AU - Serban Negru AU - Eirini Papadopoulou AU - Angela Apessos AU - Dana Lucia Stanculeanu AU - Eliade Ciuleanu AU - Constantin Volovat AU - Adina Croitoru AU - Stylianos Kakolyris AU - Gerasimos Aravantinos AU - Nikolaos Ziras AU - Elias Athanasiadis AU - Nikolaos Touroutoglou AU - Nikolaos Pavlidis AU - Haralabos P Kalofonos AU - George Nasioulas Y1 - 2014/05/01 UR - http://bmjopen.bmj.com/content/4/5/e004652.abstract N2 - Objectives Treatment decision-making in colorectal cancer is often guided by tumour tissue molecular analysis. The aim of this study was the development and validation of a high-resolution melting (HRM) method for the detection of KRAS, NRAS and BRAF mutations in Greek and Romanian patients with colorectal cancer and determination of the frequency of these mutations in the respective populations. Setting Diagnostic molecular laboratory located in Athens, Greece. Participants 2425 patients with colorectal cancer participated in the study. Primary and secondary outcome measures 2071 patients with colorectal cancer (1699 of Greek and 372 of Romanian origin) were analysed for KRAS exon 2 mutations. In addition, 354 tumours from consecutive patients (196 Greek and 161 Romanian) were subjected to full KRAS (exons 2, 3 and 4), NRAS (exons 2, 3 and 4) and BRAF (exon 15) analysis. KRAS, NRAS and BRAF mutation detection was performed by a newly designed HRM analysis protocol, followed by Sanger sequencing. Results KRAS exon 2 mutations (codons 12/13) were detected in 702 of the 1699 Greek patients with colorectal carcinoma analysed (41.3%) and in 39.2% (146/372) of the Romanian patients. Among the 354 patients who were subjected to full KRAS, NRAS and BRAF analysis, 40.96% had KRAS exon 2 mutations (codons 12/13). Among the KRAS exon 2 wild-type patients 15.31% harboured additional RAS mutations and 12.44% BRAF mutations. The newly designed HRM method used showed a higher sensitivity compared with the sequencing method. Conclusions The HRM method developed was shown to be a reliable method for KRAS, NRAS and BRAF mutation detection. Furthermore, no difference in the mutation frequency of KRAS, NRAS and BRAF was observed between Greek and Romanian patients with colorectal cancer. ER -