BACKGROUND: To compare high resolution melting (HRM) and primer extension (PE) for mutation screening of KRAS codons 12 and 13. METHODS: DNA samples were isolated from 60 colorectal cancer (CRC) patients. HRM and primer extension analyses were used for mutation screening of KRAS codons 12 and 13. RESULTS: Both methods can detect KRAS mutations in a DNA mixture containing as little as 5% mutant DNA. The concordant rate between the two methods was 100%. HRM analysis is able to distinguish mutant from wild type, however, it is unable to detect the actual base change. PE analysis needs more procedures, is time-consuming and slightly more expensive but it is able to detect the precise mutation. CONCLUSIONS: This study showed that HRM is a reliable screening method to identify mutations in the KRAS gene, and the PE can be used as a diagnostic method to accurately pinpoint the nature of mutations.
BACKGROUND: To compare high resolution melting (HRM) and primer extension (PE) for mutation screening of KRAS codons 12 and 13. METHODS: DNA samples were isolated from 60 colorectal cancer (CRC) patients. HRM and primer extension analyses were used for mutation screening of KRAS codons 12 and 13. RESULTS: Both methods can detect KRAS mutations in a DNA mixture containing as little as 5% mutant DNA. The concordant rate between the two methods was 100%. HRM analysis is able to distinguish mutant from wild type, however, it is unable to detect the actual base change. PE analysis needs more procedures, is time-consuming and slightly more expensive but it is able to detect the precise mutation. CONCLUSIONS: This study showed that HRM is a reliable screening method to identify mutations in the KRAS gene, and the PE can be used as a diagnostic method to accurately pinpoint the nature of mutations.
Authors: Giuseppe Bronte; Nicola Silvestris; Marta Castiglia; Antonio Galvano; Francesco Passiglia; Giovanni Sortino; Giuseppe Cicero; Christian Rolfo; Marc Peeters; Viviana Bazan; Daniele Fanale; Antonio Giordano; Antonio Russo Journal: Oncotarget Date: 2015-09-22