Idylla assay and next generation sequencing: an integrated EGFR mutational testing algorithm

J Clin Pathol. 2018 Aug;71(8):745-750. doi: 10.1136/jclinpath-2018-205197. Epub 2018 May 24.

Abstract

Aims: Any reference laboratory testing non-small cell lung cancer samples for predictive biomarkers needs to develop and validate a wide range of different molecular techniques, each with a specific time requirement and application. Updated international guidelines suggest that next generation sequencing (NGS) to be the initial procedure. However, in a non-negligible subset of cases, library generation may fail or amplicon coverage may be insufficient. In these NGS 'invalid' cases, the Idylla system may represent a viable option for rapid epidermal growth factor receptor (EGFR) genotyping.

Methods: This retrospective study included 68 archival DNA samples previously processed by Ion Torrent NGS assay. Out of these, 43 cases, including 24 EGFR mutant samples, had a valid NGS result, whereas 25/68 (37%) were invalid. All samples were retested by directly pipetting the DNA inside the EGFR Idylla assay cartridge.

Results: In all 43 cases with a valid NGS result, Idylla confirmed the EGFR mutational status. In particular, 24/24 (100%) of EGFR mutant samples as detected by NGS were confirmed by Idylla. Moreover, a large portion of cases (20/25; 80%) whose assessment by NGS was invalid were adequately processed by Idylla. Noteworthy, in 4/25 (16%) of cases, Idylla detected actionable EGFR mutations.

Conclusions: Idylla assay could be very useful to quickly process cases for which NGS does not allow genotyping.

Keywords: EGFR; Idylla; NGS; lung cancer; predictive biomarkers.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Algorithms*
  • Automation, Laboratory
  • Biomarkers, Tumor / genetics*
  • Carcinoma, Non-Small-Cell Lung / genetics*
  • Carcinoma, Non-Small-Cell Lung / pathology
  • DNA Mutational Analysis / instrumentation
  • DNA Mutational Analysis / methods*
  • ErbB Receptors / genetics*
  • Female
  • High-Throughput Nucleotide Sequencing* / instrumentation
  • Humans
  • Lung Neoplasms / genetics*
  • Lung Neoplasms / pathology
  • Male
  • Middle Aged
  • Mutation*
  • Predictive Value of Tests
  • Real-Time Polymerase Chain Reaction
  • Reproducibility of Results
  • Retrospective Studies

Substances

  • Biomarkers, Tumor
  • EGFR protein, human
  • ErbB Receptors