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  • Moving towards a local testing solution for undetermined thyroid fine-needle aspirates: validation of a novel custom DNA-based NGS panel

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Original research
Moving towards a local testing solution for undetermined thyroid fine-needle aspirates: validation of a novel custom DNA-based NGS panel
  1. Roberta Sgariglia1,
  2. Mariantonia Nacchio1,
  3. Ilaria Migliatico1,
  4. Elena Vigliar1,
  5. Umberto Malapelle1,
  6. Pasquale Pisapia1,
  7. Caterina De Luca1,
  8. Antonino Iaccarino1,
  9. Domenico Salvatore1,
  10. Stefania Masone2,
  11. Giancarlo Troncone1,
  12. Claudio Bellevicine1
  1. 1 Public Health, University of Naples Federico II, Naples, Italy
  2. 2 Clinical Medicine and Surgery, General Surgery Unit, University of Naples Federico II, Naples, Italy
  1. Correspondence to Professor Giancarlo Troncone, Public Health, University of Naples Federico II, 80131 Naples, Italy; giancarlo.troncone{at}unina.it

Abstract

Aims In thyroid cytopathology, the undetermined diagnostic categories still pose diagnostic challenges. Although next-generation sequencing (NGS) is a promising technique for the molecular testing of thyroid fine-needle aspiration (FNA) specimens, access to such technology can be difficult because of its prohibitive cost and lack of reimbursement in countries with universal health coverage. To overcome these issues, we developed and validated a novel custom NGS panel, Nexthyro, specifically designed to target 264 clinically relevant mutations involved in thyroid tumourigenesis. Moreover, in this study, we compared its analytical performance with that of our previous molecular testing strategy.

Methods The panel, which includes 15 genes (BRAF, EIF1AX, GNAS, HRAS, IDH1, KRAS, NF2, NRAS, PIK3CA, PPM1D, PTEN, RET, DICER1, CHEK2, TERT promoter), was validated with a cell-line derived reference standard and 72 FNA archival samples previously tested with the 7-gene test.

Results Nexthyro yielded 100% specificity and detected mutant alleles at levels as low as 2%. Moreover, in 5/72 (7%) FNAs, it detected more clinically relevant mutations in BRAF and RAS genes compared with the 7-gene test. Nexthyro also revealed better postsequencing metrics than the previously adopted commercial ‘generic’ NGS panel.

Conclusion Our comparative analysis indicates that Nexthyro is a reliable NGS panel. The study also implies that a custom-based solution for routine thyroid FNA is sustainable at the local level, allowing patients with undetermined thyroid nodules affordable access to NGS.

  • thyroid neoplasms
  • cytological techniques
  • pathology
  • molecular

Data availability statement

All data relevant to the study are included in the article or uploaded as online supplemental information.

https://doi.org/10.1136/jclinpath-2021-207429

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    Data availability statement

    All data relevant to the study are included in the article or uploaded as online supplemental information.

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    Footnotes

    • Handling editor Runjan Chetty.

    • Twitter @UmbertoMalapel1, @PasqualePisapia

    • RS and MN contributed equally.

    • Contributors Conceptualisation, RS, MN, GT and CB; methodology, all authors; software, all authors; validation, all authors; formal analysis, all authors; investigation, all authors; resources, all authors; data curation, all authors; writing—original draft preparation, RS, MN, CB; writing—review and editing, all authors; visualisation, all authors; supervision, GT and CB; project administration, RS, MN, GT and CB; funding acquisition, GT and CB.

    • Funding Regione Campania for the investigation of the molecular biology of thyroid cancer. Grant number: LR n.24 29/12/2005.

    • Competing interests EV has received personal fees (as consultant and/or speaker bureau) from Diaceutics, unrelated to the current work. UM has received personal fees (as consultant and/or speaker bureau) from Boehringer Ingelheim, Roche, MSD, Amgen, Thermo Fisher Scientifics, Diaceutics, GSK, Merck and AstraZeneca, unrelated to the current work. GT reports personal fees (as speaker bureau or advisor) from Roche, MSD, Pfizer and Bayer, unrelated to the current work.

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

    • Supplemental material This content has been supplied by the author(s). It has not been vetted by BMJ Publishing Group Limited (BMJ) and may not have been peer-reviewed. Any opinions or recommendations discussed are solely those of the author(s) and are not endorsed by BMJ. BMJ disclaims all liability and responsibility arising from any reliance placed on the content. Where the content includes any translated material, BMJ does not warrant the accuracy and reliability of the translations (including but not limited to local regulations, clinical guidelines, terminology, drug names and drug dosages), and is not responsible for any error and/or omissions arising from translation and adaptation or otherwise.