Article Text

Download PDFPDF
Comparison of two next-generation sequencing-based approaches for liquid biopsy analysis in patients with non-small cell lung cancer: a multicentre study
  1. Silvia Bessi1,
  2. Francesco Pepe2,
  3. Gianluca Russo2,
  4. Pasquale Pisapia2,
  5. Marco Ottaviantonio1,
  6. Francesca Biancalani2,
  7. Antonino Iaccarino2,
  8. Maria Russo2,
  9. Mauro Biancalani3,
  10. Giancarlo Troncone2,
  11. Umberto Malapelle2
  1. 1 Departmental Structure of Oncological Molecular Pathology, Azienda USL Toscana Centro, Prato, Italy
  2. 2 Public Health, University of Naples Federico II, Naples, Italy
  3. 3 Morphological Diagnostic and Biomolecular Characterization Area, Complex Unit of Pathological Anatomy, Azienda USL Toscana Centro, Prato, Italy
  1. Correspondence to Professor Giancarlo Troncone, Public Health, University of Naples Federico II, Naples, Italy; giancarlo.troncone{at}


In the era of personalised medicine, testing for an increasing number of predictive biomarkers is becoming a priority. However, tissue biopsies from these patients are oftentimes insufficient for conventional approaches, a common issue that deprives them of the clinical benefits of biomarker-directed treatments. To tackle this problem, many clinical laboratories are resorting to circulating tumour DNA (ctDNA), which is becoming increasingly appreciated as a valuable source for biomarker testing. In this context, next-generation sequencing (NGS) has become essential. Indeed, different NGS systems are able to detect several clinically relevant low-frequency hot-spot mutations simultaneously in a single run. However, their reproducibility in the analysis of ctDNA has not yet been investigated. The purpose of this study was to evaluate the reproducibility of using Illumina MiSeq and Thermo Fisher Ion S5 Plus platforms to assess pathogenic alterations in non-small cell lung cancer (NSCLC) liquid biopsy specimens. Using the in vitro diagnostic (IVD) NGS panel Myriapod NGS Cancer panel DNA (Diatech Pharmacogenetics) on MiSeq platform (Illumina), we reanalysed ctDNA extracted from a retrospective series of n=40 patients with advanced NSCLC previously tested with a custom NGS panel (SiRe) on Thermo Fisher Ion S5 Plus system. Overall, 13 out of 40 (32.5%) ctDNA samples displayed pathogenic alterations in at least two genes, namely, EGFR and KRAS. A concordance rate of 100% was identified between the two methodologies in terms of sample mutational status and total number of detected variables. All NGS platforms featured a high degree of concordance.

  • Pathology, Molecular
  • Lung Neoplasms
  • LUNG

Statistics from

Request Permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.


  • SB and FP are joint first authors.

  • Handling editor Runjan Chetty.

  • Twitter @PasqualePisapia, @UmbertoMalapel1

  • SB and FP contributed equally.

  • Contributors Conceptualisation: UM; writing—original draft preparation: GR, FP, PP and UM; supervision and project administration: SB, GT and UM; funding acquisition: GT; methodology, software, validation, formal analysis, investigation, resources, data curation, writing—review and editing and visualisation: all authors.

  • Funding Monitoraggio ambientale, studio ed approfondimento della salute della popolazione residente in aree a rischio—In attuazione della D.G.R. Campanian.180/2019. POR Campania FESR 2014–2020 Progetto 'Sviluppo di Approcci Terapeutici Innovativi per patologie Neoplastiche resistenti ai trattamenti—SATIN'.

  • Competing interests PP received personal fees as a speaker bureau of Novartis, unrelated to the current work. GT reported personal fees (as member of the speaker bureau or advisor) from Roche, MSD, Pfizer, Boehringer Ingelheim, Eli Lilly, BMS, GSK, Menarini, AstraZeneca, Amgen and Bayer, unrelated to the current work. UM received personal fees (as consultant and/or speaker bureau) from Boehringer Ingelheim, Roche, MSD, Amgen, Thermo Fisher Scientifics, Eli Lilly, Diaceutics, GSK, Merck and AstraZeneca, Janssen, Diatech, Novartis and Hedera, unrelated to the current work.

  • Provenance and peer review Not commissioned; externally peer reviewed.