Article Text


Multicentre validation of a microRNA-based assay for diagnosing indeterminate thyroid nodules utilising fine needle aspirate smears
  1. Gila Lithwick-Yanai1,
  2. Nir Dromi1,
  3. Alexander Shtabsky2,3,
  4. Sara Morgenstern3,4,
  5. Yulia Strenov3,4,
  6. Meora Feinmesser3,4,
  7. Vladimir Kravtsov3,5,
  8. Marino E Leon6,
  9. Marián Hajdúch7,
  10. Syed Z Ali8,
  11. Christopher J VandenBussche8,
  12. Xinmin Zhang9,10,
  13. Leonor Leider-Trejo2,3,
  14. Asia Zubkov2,
  15. Sergey Vorobyov11,
  16. Michal Kushnir1,
  17. Yaron Goren1,12,
  18. Sarit Tabak1,
  19. Etti Kadosh1,
  20. Hila Benjamin13,
  21. Temima Schnitzer-Perlman1,
  22. Hagai Marmor1,
  23. Maria Motin1,
  24. Danit Lebanony1,
  25. Sharon Kredo-Russo1,
  26. Heather Mitchell13,
  27. Melissa Noller13,
  28. Alexis Smith13,
  29. Olivia Dattner13,
  30. Karin Ashkenazi13,
  31. Mats Sanden13,
  32. Kenneth A Berlin13,
  33. Dganit Bar1,
  34. Eti Meiri1
  1. 1Rosetta Genomics Ltd, Rehovot, Israel
  2. 2Pathology Institute, Tel-Aviv Sourasky Medical Center, Tel Aviv, Israel
  3. 3The Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel
  4. 4Pathology Institute, Rabin Medical Center, Petach Tikva, Israel
  5. 5Pathology Institute, Meir Medical Center, Kfar Saba, Israel
  6. 6H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, USA
  7. 7Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic
  8. 8The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
  9. 9Temple University Hospital, Philadelphia, Pennsylvania, USA
  10. 10Cooper University Hospital, Cooper Medical School of Rowan University at Camden, New Jersey, USA
  11. 11National Centre of Clinical and Morphological Diagnostics, St Petersburg, Russia
  12. 12Geha Mental Health Center, Petach Tikva, Israel
  13. 13Rosetta Genomics Inc, Philadelphia, Pennsylvania, USA
  1. Correspondence to Dr Eti Meiri, Rosetta Genomics Ltd, 10 Plaut St, Rehovot 7670609, Israel; meirie{at}


Aims The distinction between benign and malignant thyroid nodules has important therapeutic implications. Our objective was to develop an assay that could classify indeterminate thyroid nodules as benign or suspicious, using routinely prepared fine needle aspirate (FNA) cytology smears.

Methods A training set of 375 FNA smears was used to develop the microRNA-based assay, which was validated using a blinded, multicentre, retrospective cohort of 201 smears. Final diagnosis of the validation samples was determined based on corresponding surgical specimens, reviewed by the contributing institute pathologist and two independent pathologists. Validation samples were from adult patients (≥18 years) with nodule size >0.5 cm, and a final diagnosis confirmed by at least one of the two blinded, independent pathologists. The developed assay, RosettaGX Reveal, differentiates benign from malignant thyroid nodules, using quantitative RT-PCR.

Results Test performance on the 189 samples that passed quality control: negative predictive value: 91% (95% CI 84% to 96%); sensitivity: 85% (CI 74% to 93%); specificity: 72% (CI 63% to 79%). Performance for cases in which all three reviewing pathologists were in agreement regarding the final diagnosis (n=150): negative predictive value: 99% (CI 94% to 100%); sensitivity: 98% (CI 87% to 100%); specificity: 78% (CI 69% to 85%).

Conclusions A novel assay utilising microRNA expression in cytology smears was developed. The assay distinguishes benign from malignant thyroid nodules using a single FNA stained smear, and does not require fresh tissue or special collection and shipment conditions. This assay offers a valuable tool for the preoperative classification of thyroid samples with indeterminate cytology.


This is an Open Access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:

Statistics from

Review history and Supplementary material

  • Abstract in Hebrew

    This web only file has been produced by the BMJ Publishing Group from an electronic file supplied by the author(s) and has not been edited for content.


  • GL-Y and ND contributed equally.

  • Handling editor Runjan Chetty

  • Contributors GL-Y analysed the data, developed the algorithm and the software, and wrote the manuscript. ND, TS-P, HB analysed the data, developed the algorithm and the software, and reviewed and edited the manuscript. ASh collected data, reviewed the slides, was involved in interpretation of the data, and reviewed and edited the manuscript. SM, YS, MF, VK, MEL, MH, SZA, CJVB, XZ, AZ, SV identified clinical cases, reviewed the slides and collected data. LL-T and MS reviewed and analysed slides. MK contributed to experimental design, collected the data, was involved in interpretation of the data, and reviewed and edited the manuscript. YG contributed to experimental design and collected the data. ST, EK, HM, MM, DL, SK-R, HM, MN, ASm, OD, KA performed experiments and were involved in calibrations and protocol development. KAB was involved in conceptualisation and reviewed and edited the manuscript. DB was involved in conceptualisation, was involved in interpretation of the data, and wrote the manuscript. EM was involved in conceptualisation, overviewed experimental design and procedures, was involved in interpretation of the data, and reviewed and edited the manuscript. All authors read and approved the final manuscript.

  • Competing interests Authors affiliated with Rosetta Genomics are full-time employees of the company and/or hold equity in the company, which stands to gain from the publication of this manuscript. One of the authors (A. Shtabsky) is a payed consultant for Rosetta Genomics. The authors from medical/clinical centres have received research funding from the company as part of this and/or other collaborative projects.

  • Ethics approval IRB (for each institution).

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

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.