Utility of Select Gene Mutation Detection in Tumors by the Idylla Rapid Multiplex PCR Platform in Comparison to Next-Generation Sequencing

Genes (Basel). 2022 Apr 29;13(5):799. doi: 10.3390/genes13050799.


Testing of tumors by next generation sequencing (NGS) is impacted by relatively long turnaround times and a need for highly trained personnel. Recently, Idylla oncology assays were introduced to test for BRAF, EGFR, KRAS, and NRAS common hotspot mutations that do not require specialized trained personnel. Moreover, the interpretation of results is fully automated, with rapid turnaround time. Though Idylla testing and NGS have been shown to have high concordance in identifying EGFR, BRAF, KRAS, and NRAS hotspot mutations, there is limited experience on optimal ways the Idylla system can be used in routine practice. We retrospectively evaluated all cases with EGFR, BRAF, KRAS, or NRAS mutations identified in clinical specimens sequenced on two different NGS panels at the University of Rochester Medical Center (URMC) molecular diagnostics laboratory between July 2020 and July 2021 and assessed if these mutations would be detected by the Idylla cartridges if used. We found that the Idylla system could accurately identify Tier 1 or 2 actionable genomic alterations in select associated disease pathologies if used. Yet, in a minority of cases, we would have been unable to detect NGS-identified pathogenic mutations due to their absence on the Idylla panels. We derived algorithmic practice guidelines for the use of the Idylla cartridges. Overall, Idylla molecular testing could be implemented either as a first-line standalone diagnostic tool in select indications or for orthogonal confirmation of uncertain results.

Keywords: Idylla platform; molecular diagnostics; next generation sequencing.

MeSH terms

  • DNA Mutational Analysis / methods
  • ErbB Receptors / genetics
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Multiplex Polymerase Chain Reaction
  • Mutation
  • Neoplasms* / diagnosis
  • Neoplasms* / genetics
  • Proto-Oncogene Proteins B-raf* / genetics
  • Proto-Oncogene Proteins p21(ras) / genetics
  • Retrospective Studies


  • ErbB Receptors
  • Proto-Oncogene Proteins B-raf
  • Proto-Oncogene Proteins p21(ras)

Grant support

This research received no external funding.