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

J Clin Pathol. 2017 Jul;70(7):610-614. doi: 10.1136/jclinpath-2017-204356. Epub 2017 Mar 14.

Abstract

Background: Molecular testing is increasingly needed in colorectal carcinoma (CRC) and the current clinically relevant mutations are in BRAF, KRAS and NRAS. 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. BRAF IHC showed 90% concordance with NGS (95% CI 70% to 97%).

Conclusions: This study validates Idylla in formalin-fixed, paraffin-embedded CRC tissue. BRAF IHC, however, is an unreliable substitute for molecular testing in CRC.

Keywords: COLORECTAL CANCER; IMMUNOHISTOCHEMISTRY; MOLECULAR PATHOLOGY.

Publication types

  • Comparative Study
  • Validation Study

MeSH terms

  • Colorectal Neoplasms / diagnosis*
  • Colorectal Neoplasms / genetics
  • GTP Phosphohydrolases / genetics
  • Humans
  • Immunohistochemistry
  • Membrane Proteins / genetics
  • Mutation / genetics
  • Point-of-Care Systems
  • Proto-Oncogene Proteins B-raf / genetics
  • Proto-Oncogene Proteins p21(ras) / genetics
  • Real-Time Polymerase Chain Reaction / methods

Substances

  • KRAS protein, human
  • Membrane Proteins
  • Proto-Oncogene Proteins B-raf
  • GTP Phosphohydrolases
  • NRAS protein, human
  • Proto-Oncogene Proteins p21(ras)