eprintid: 19380
rev_number: 38
eprint_status: archive
userid: 1589
dir: disk0/00/01/93/80
datestamp: 2015-12-09 09:33:41
lastmod: 2024-04-17 21:20:21
status_changed: 2015-12-09 09:33:41
type: article
metadata_visibility: show
creators_name: Kriegsmann, Mark
creators_name: Arens, Norbert
creators_name: Endris, Volker
creators_name: Weichert, Wilko
creators_name: Kriegsmann, Jörg
title: Detection of KRAS, NRAS and BRAF by mass spectrometry - a sensitive, reliable, fast and cost-effective technique
subjects: ddc-610
divisions: i-912000
abstract: 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.  
date: 2015
publisher: BioMed Central
id_scheme: DOI
ppn_swb: 1656046091
own_urn: urn:nbn:de:bsz:16-heidok-193805
language: eng
bibsort: KRIEGSMANNDETECTIONO2015
full_text_status: public
publication: Diagnostic pathology
volume: 10
number: 132
place_of_pub: London
pagerange: 1-11
issn: 1746-1596
citation:   Kriegsmann, Mark ; Arens, Norbert ; Endris, Volker ; Weichert, Wilko ; Kriegsmann, Jörg  (2015) Detection of KRAS, NRAS and BRAF by mass spectrometry - a sensitive, reliable, fast and cost-effective technique.  Diagnostic pathology, 10 (132).  pp. 1-11.  ISSN 1746-1596     
document_url: https://archiv.ub.uni-heidelberg.de/volltextserver/19380/1/13000_2015_Article_364.pdf