%0 Journal Article
%@ 1746-1596
%A Kriegsmann, Mark
%A Arens, Norbert
%A Endris, Volker
%A Weichert, Wilko
%A Kriegsmann, Jörg
%C London
%D 2015
%F heidok:19380
%I BioMed Central
%J Diagnostic pathology
%N 132
%P 1-11
%T Detection of KRAS, NRAS and BRAF by mass spectrometry - a sensitive, reliable, fast and cost-effective technique
%U https://archiv.ub.uni-heidelberg.de/volltextserver/19380/
%V 10
%X Background: According to current clinical guidelines mutational analysis for KRAS and NRAS is recommended prior to EGFR-directed therapy of colorectal cancer (CRC) in the metastatic setting. Therefore, reliable, fast, sensitive and cost-effective methods for routine tissue based molecular diagnostics are required that allow the assessment of the CRC mutational status in a high throughput fashion.   Methods: We have developed a custom designed assay for routine mass-spectrometric (MS) (MassARRAY®, Agena Bioscience) analysis to test the presence/absence of 18 KRAS, 14 NRAS and 4 BRAF mutations. We have applied this assay to 93 samples from patients with CRC and have compared the results with Sanger sequencing and a chip hybridization assay (KRAS LCD-array Kit, Chipron). In cases with discordant results, next-generation sequencing (NGS) was performed.   Results: MS detected a KRAS mutation in 46/93 (49 %), a NRAS mutation in 2/93 (2 %) and a BRAF mutation in 1/93 (1 %) of the cases. MS results were in agreement with results obtained by combination of the two other methods in 92 (99 %) of 93 cases. In 1/93 (1 %) of the cases a G12V mutation has been detected by Sanger sequencing and MS, but not by the chip assay. In this case, NGS has confirmed the G12V mutation in KRAS.   Conclusions: Mutational analysis by MS is a reliable method for routine diagnostic use, which can be easily extended for testing of additional mutations.