Article Text

other Versions

PDF
Sanger sequencing in routine KRAS testing: a review of 1720 cases from a pathologist's perspective
  1. Umberto Malapelle1,
  2. Claudio Bellevicine1,
  3. Maria Salatiello1,
  4. Caterina de Luca1,
  5. Elisabetta Rispo1,
  6. Palmira Riccio2,
  7. Lucianna Sparano2,
  8. Alfonso De Stefano3,
  9. Chiara Carlomagno3,
  10. Francesco Maria Maiello4,
  11. Giulia Vita5,
  12. Oscar Nappi6,
  13. Giancarlo Troncone1
  1. 1Scienze Biomorfologiche e Funzionali, Università degli Studi di Napoli Federico II, Napoli, Italy
  2. 2Anatomia Patologica, Azienda Ospedaliera Vincenzo Scarlato, Salerno, Italy
  3. 3Endocrinologia ed Oncologia Molecolare e Clinica, Università degli Studi di Napoli Federico II, Napoli, Italy
  4. 4Anatomia Patologica, Ospedale dei Pellegrini, Napoli, Italy
  5. 5Anatomic Pathology, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Centro di Riferimento Oncologico della Basilicata (CROB), Rionero in Vulture, Potenza, Italy
  6. 6Anatomia Patologica, AORN Antonio Cardarelli, Napoli, Italy
  1. Correspondence to Professor Giancarlo Troncone, Scienze Biomorfologiche e Funzionali, University of Naplese Federico II, via Sergio Pansini 5, Napoli 80128, Italy; giancarlo.troncone{at}unina.it

Abstract

Background Sanger sequencing (SS) of PCR products is still the most frequent method to test colorectal cancer for KRAS mutations in routine practice.

Methods An audit of SS on 1720 routine cases was carried out, taking into account age, gender, specimen type (resection vs biopsies), tumour site (primary vs metastasis), tumour stage, neoplastic cells abundance (>30% vs <30%) and fixation type (buffered formalin vs simple formalin). In a subset of 50 wild-type (WT) patients correlations between SS findings and response rate (RR), progression-free survival (PFS) and overall survival (OS) were also evaluated.

Results The tests were informative in 1691 cases (98.3%). Mutations were detected in 671 cases (39.6%). No significant differences in mutation rates were observed with respect to age (p=0.2), gender (p=0.2), specimen type (p=0.3) and formalin fixation (p=0.08). Conversely, KRAS mutant rate was higher in metastatic tissue (50% vs 39%, p=0.02), in samples with over 30% of neoplastic cells (43.4% vs 26.6%, p=0.02) and in tumours tested in stage IV (p=0.05). The RR of SS KRAS WT patients was 26% (one complete and 12 partial responses). The disease control rate (objective responses plus stable disease) was 56%. Median PFS was 4.4 months and median OS was 10.4 months.

Conclusions Pathological criteria that make SS a more robust method for KRAS testing and treatment response prediction are neoplastic cell abundance, metastatic tissue sample and stage IV primary tumour.

  • Colorectal Cancer
  • Audit
  • Histopathology
  • Laboratory Tests
  • Molecular Pathology

Statistics from Altmetric.com

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.