gms | German Medical Science

27. Deutscher Krebskongress

Deutsche Krebsgesellschaft e. V.

22. - 26.03.2006, Berlin

Disseminated tumor cells in bone marrow of breast cancer patients - Comparison of immunocytochemistry and RT-PCR

Meeting Abstract

  • corresponding author presenting/speaker Malgorzata Banys - Universitäts-Frauenklinik Tübingen, Tuebingen, Deutschland
  • Sven Becker - Universitäts-Frauenklinik Tübingen
  • Silke Dürr-Störzer - Universitäts-Frauenklinik Tübingen
  • Graziella Becker-Pergola - Universitäts-Frauenklinik Tübingen
  • Natalia Krawczyk - Universitäts-Frauenklinik Tübingen
  • Diethelm Wallwiener - Universitäts-Frauenklinik Tübingen
  • Erich-Franz Solomayer - Universitäts-Frauenklinik Tübingen
  • Tanja Fehm - Universitäts-Frauenklinik Tübingen

27. Deutscher Krebskongress. Berlin, 22.-26.03.2006. Düsseldorf, Köln: German Medical Science; 2006. DocPE099

Die elektronische Version dieses Artikels ist vollständig und ist verfügbar unter:

Veröffentlicht: 20. März 2006

© 2006 Banys et al.
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Background: Since 1999 immunocytochemistry (ICC) has been regarded as the standard method of choice for detection of disseminated tumor cells (DTC) in bone marrow (BM). In the meantime molecular-based assays were developed, such as RT-PCR. The purpose of our investigation was to evaluate both techniques with respect to their sensitivities on the basis of 299 bone marrow aspirates from breast cancer patients.

Methods: 299 bone marrow aspirates from primary breast cancer patients were processed with both methods.

Immunocytochemistry: After Ficoll enrichment of 10ml bone marrow, cytospins were prepared and stained using the A45-B/B3 primary antibody for pCK. Cytospins were analyzed using the ACIS system (Chromavision) according to the ISHAGE evaluation criteria.

RT-PCR: mRNA was extracted and purified using the mRNA isolation for blood and bone marrow kit (Roche® Molecular Biochemicals). RT-PCR was performed on the LightCycler® system, using the RNAMaster Hybridization Probes kit and custom primers and probes. Primers were selected to amplify a 380bp fragment of the CK19 gene.

Positive and negative controls were included with each batch of samples for both procedures.

Results: Disseminated tumor cells could be detected in 53% (157 out of 299) of the aspirates by at least one of the methods. The positivity rates of ICC and RT-PCR were 37% and 39%, respectively. The concordance rate between ICC and RT-PCR was 71%. Discordant results were found in 87 patients.

46 BM aspirates were positive by RT-PCR but negative by ICC whereas 41 samples were positive only by ICC.

Conclusions: Both methods have their advantages. ICC allows an interpretation based on morphological criteria and is a standardized procedure with well-known correlation to the prognosis but it is observer-dependent and labor intensive. RT-PCR is time-efficient and may increase the sensitivity but it lacks a standard protocol. We conclude that RT-PCR-based assays have a potential to improve diagnostics in this field. Further collaborative efforts must be made to establish a standard protocol for RT-PCR for the detection of disseminated tumor cells in bone marrow.