title: Validating a fully automated real-time PCR-based system for use in the molecular diagnostic analysis of colorectal carcinoma: a comparison with NGS and IHC.
abstract: BACKGROUND
  Molecular testing is increasingly needed in colorectal carcinoma (CRC) and the current clinically relevant mutations are inBRAF,KRASandNRAS. This study aimed to further validate a new alternative polymerase chain reaction (PCR) platform (Idylla, Biocartis) against existing next-generation sequencing (NGS) and immunohistochemistry (IHC) assays.
  
  
  METHODS
  56 Idylla tests were performed on 43 CRC cases, in a total of 74 comparisons against an NGS panel (Ion Torrent) and the VE1 (anti-BRAF) antibody IHC. Discrepant cases were also compared with either conventional (Cobas) or droplet digital PCR (Bio-Rad).
  
  
  RESULTS
  Idylla showed an overall concordance of 100% (95% CI 93% to 100%) with comparator molecular testing and indications were that Idylla is likely to be more sensitive than routine NGS.BRAFIHC showed 90% concordance with NGS (95% CI 70% to 97%).
  
  
  CONCLUSIONS
  This study validates Idylla in formalin-fixed, paraffin-embedded CRC tissue.BRAFIHC, however, is an unreliable substitute for molecular testing in CRC.
authors.affiliation: Department of Cellular Pathology, Oxford University Hospitals NHS Trust, John Radcliffe Hospital, Oxford, UK.|Oxford Molecular Diagnostics Centre, Molecular Haematology, Oxford University Hospitals NHS Trust, John Radcliffe Hospital, Oxford, UK.|Department of Oncology, University of Oxford, John Radcliffe Hospital, Oxford, UK.
authors.forename: Richard
authors.initials: R
authors.lastname: Colling
authors.affiliation: Department of Cellular Pathology, Oxford University Hospitals NHS Trust, John Radcliffe Hospital, Oxford, UK.|Department of Laboratory Medicine, Changi General Hospital, Singapore, Singapore.|Ludwig Institute for Cancer Research, University of Oxford, Old Road Campus Research Building, Oxford, UK.
authors.forename: Lai Mun
authors.initials: LM
authors.lastname: Wang
authors.affiliation: Department of Cellular Pathology, Oxford University Hospitals NHS Trust, John Radcliffe Hospital, Oxford, UK.|Nuffield Division of Clinical Laboratory Sciences, University of Oxford, John Radcliffe Hospital, Oxford, UK.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Soilleux
mesh_terms: D015179:Colorectal Neoplasms / Q000175:diagnosis* / Q000235:genetics; D020558:GTP Phosphohydrolases / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000235:genetics; D009154:Mutation / Q000235:genetics; D019095:Point-of-Care Systems; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
chemical_list: C117307:KRAS protein, human; D008565:Membrane Proteins; D048493:Proto-Oncogene Proteins B-raf; D020558:GTP Phosphohydrolases; C579846:NRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)
keywords: COLORECTAL CANCER; IMMUNOHISTOCHEMISTRY; MOLECULAR PATHOLOGY