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How to prepare cytological samples for molecular testing
  1. Claudio Bellevicine1,
  2. Umberto Malapelle1,
  3. Elena Vigliar1,
  4. Pasquale Pisapia1,
  5. Giulia Vita2,
  6. Giancarlo Troncone1
  1. 1Department of Public Health, Anatomic Pathology Unit, University of Napoli Federico II School of Medicine, Napoli, Italy
  2. 2Anatomia Patologica, I.R.C.S. CROB, Rionero in Vulture, Potenza, Italy
  1. Correspondence to Professor Giancarlo Troncone, Department of Public Health, University of Naples Federico II, via Sergio Pansini 5, Naples I/80131, Italy; giancarlo.troncone{at}


This review is focused on the challenges in standardising and optimising molecular testing workflow in cytopathology. Although cytological samples yield optimal quality DNA, whose minimal amounts in most cases suffice even for multigene mutational profiling, the success of molecular testing is strongly dependent on standardised preanalytical protocols for maximising DNA yield and quality. Sample cytopreparation influences, even more, the quality of RNA and consequently the potential success of reverse transcription-PCR. Here, the educational and technical involvement of the cytopathologist as a relevant component of a multidisciplinary team, in the issues related to test request, specimen collection, fixation, processing, staining, tumour fraction enrichment, DNA quality/quantity assessment and storage conditions is discussed. In addition, the specific sample requirements related to more recent technological developments are examined, underlining the modern role of the cytopathologist, whose continuous education is crucial to meet the opportunities of molecular medicine.

  • personalized medicine
  • cancer
  • cytopathology
  • fine-needle aspiration
  • molecular cytopathology
  • polymerase chain reaction
  • next generation sequencing
  • mutational analysis
  • preanalytic
  • direct smear
  • cell block

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  • Handling editor Runjan Chetty.

  • Contributors CB and GT conceived the review and wrote the paper; CB, UM, EV, PP, GV and GT collected and assembled the literature data.

  • Competing interests None declared.

  • Patient consent Obtained.

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