KRAS MUTATION TESTING ON Q3 SYSTEM

Background: Colon Rectal Cancer (CRC) is an aggressive human malignancy with a complex genomic landscape harboring KRAS mutations. Activating mutations in KRAS are strongly associated with prognosis, surgical strategy and poor response to anti-EGFR therapies. To this reason, testing for KRAS mutations has become increasingly common in clinical practice to better patients management. Several methods have been employed to KRAS testing that differ with regard to their effectiveness, risk and degree of invasiveness. To provide a rapid, accurate and easy to use diagnostic test, the biosensor technologies based on genomic approaches, represent an attractive promise to offer new options in the field of screening and biomedicine. Here we propose a new method to detect simultaneously the main KRAS mutations on portable Q3 platform (STMicroelectronics) based on Real Time PCR methodology. Based on hybrid silicon-plastic technology, this miniaturized chip offers a rapid and sample-in answer-out analysis allowing a new approach to genetic counselling and testing for several diseases. Materials and Methods: The developed test for detection of KRAS includes the eight common hotspot spanning in codons 12 and 13 of exon 2. On the Q3 platform, each wells host 6 µl of total volume reactions. The sensitivity, selectivity and specificity of the Q3 were compared in a commercial instrument. Results: The Q3 chip was able to identify the mutations in KRAS that are predictive of prognosis of CRC and individual response to therapy. The performance was comparable in terms of threshold cycles (Cts) and amplification plots to those of other commercial instruments. Discussions: Data obtained revealed that miniaturized Q3 system is a rapid and efficient platform for genetic screening and its versatility could be useful for the realization of customizable genetic tests.