Intra-operative rapid diagnostic method based on CK19 mRNA expression for the detection of lymph node metastases in breast cancer

Mike Visser, Mehdi Jiwa, Anja Horstman, Antoinette A.T.P. Brink, Rene P. Pol, Paul Van Diest, Peter J.F. Snijders, Chris J.L.M. Meijer

Research output: Contribution to journalArticle

Abstract

Staging by sentinel node (SN) biopsy is the standard procedure for clinically node-negative breast cancer patients. Intra-operative analysis of the SN allows immediate axillary lymph node (ALN) dissection in SN positive patients, but a quick, reliable and reproducible method is lacking. We tested the suitability of a quantitative cytokeratin 19 (CK19) mRNA one step nucleic acid amplification (OSNA#) technique (OSNA-CK19) for intra-operative SN analysis. OSNA-CK19 involves a short manual sample preparation step and subsequent fully automated amplification of CK19 mRNA based on reverse transcription loop-mediated isothermal amplification, with results available within 30-40 min. OSNA-CK19 was compared to histological staining (Hematoxylin&Eosin and CAM5.2 and CK19 immunostaining) of 346 frozen ALNs from 32 breast cancer patients, using half of the lymph node for each method. 267 samples were negative and 61 positive by both methods. Three samples were histology positive and OSNA-CK19 negative. Fifteen samples were histology negative and OSNA-CK19 positive, 11 of which had copy numbers close to the cut-off level of OSNA-CK19. Seven of these 15 samples were RT-PCR positive for epithelial markers and/or showed CK19 protein expression by Western blot suggesting the presence of tumor deposits in the lymph node part investigated by OSNA-CK19. Concordance with histology was 94.8%, and 96.8% after exclusion of the latter 7 discordant cases. Sensitivity was 95.3% and specificity was 94.7% before and 97.1% after discordant case investigation. Our results indicate that OSNA-CK19 can potentially be useful in an intra-operative clinical setting to detect SN tumor involvement in breast cancer patients.

Original languageEnglish (US)
Pages (from-to)2562-2567
Number of pages6
JournalInternational Journal of Cancer
Volume122
Issue number11
DOIs
StatePublished - Jun 1 2008

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Keywords

  • Breast cancer
  • Cytokeratin 19
  • Intra-operative
  • Lymph node metastases
  • OSNA

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Visser, M., Jiwa, M., Horstman, A., Brink, A. A. T. P., Pol, R. P., Van Diest, P., Snijders, P. J. F., & Meijer, C. J. L. M. (2008). Intra-operative rapid diagnostic method based on CK19 mRNA expression for the detection of lymph node metastases in breast cancer. International Journal of Cancer, 122(11), 2562-2567. https://doi.org/10.1002/ijc.23451