Annals of Surgical Oncology

, Volume 12, Issue 9, pp 753–760

The Detection of Isolated Tumor Cells in Bone Marrow Comparing Bright-Field Immunocytochemistry and Multicolor Immunofluorescence

Authors

    • Department of SurgeryUniversity of Vermont College of Medicine
  • Roberto Kusminsky
    • Charleston Area Medical Center Institute
  • Edward Manna
    • Department of SurgeryUniversity of Vermont College of Medicine
  • Abiy Ambaye
    • Department of Surgical PathologyFletcher Allen Health Care
  • Donald L. Weaver
    • Department of Surgical PathologyFletcher Allen Health Care
  • Seth P. Harlow
    • Department of SurgeryUniversity of Vermont College of Medicine
  • Michael Covelli
    • Charleston Area Medical Center Institute
  • Mary A. Stanley
    • Department of SurgeryUniversity of Vermont College of Medicine
  • Laurence McCahill
    • Department of SurgeryUniversity of Vermont College of Medicine
  • Frank Ittleman
    • Department of SurgeryUniversity of Vermont College of Medicine
  • Bruce Leavitt
    • Department of SurgeryUniversity of Vermont College of Medicine
  • Martin Krag
    • Department of Orthopaedics and RehabilitationUniversity of Vermont College of Medicine
Article

DOI: 10.1245/ASO.2005.12.004

Cite this article as:
Krag, D.N., Kusminsky, R., Manna, E. et al. Ann Surg Oncol (2005) 12: 753. doi:10.1245/ASO.2005.12.004

Abstract

Background

The detection of isolated tumor cells in bone marrow by immunocytochemistry (ICC) has been reported to predict progression of early-stage breast cancer. The most common staining procedure uses bright-field ICC with cytokeratin (CK) antibodies to label isolated tumor cells. However, this method can result in false-positive staining events. We used multicolor immunofluorescence (IF) to develop a more specific assay for detecting isolated tumor cells in marrow samples from breast cancer patients.

Methods

We compared ICC and IF side by side for detection of cancer cells and false-positive staining events on bone marrow aspirates from breast cancer patients, bone marrow from healthy donors, and healthy donor blood spiked with cancer cells. The primary target for isolated tumor cell detection was CK for both methods. IF used an additional set of antibodies to label hematopoietic cells (HCs).

Results

The detection rate of CK+ events in breast cancer patient bone marrow aspirates was 18 (58%) of 31 for ICC and 21 (68%) of 31 for IF. However, with IF, 17 of 21 CK+ cases were stained with HC markers and thus were identified as false-positive events. A surprisingly high CK+ event rate was observed in healthy donor blood and marrow. In all healthy donor samples, CK+ events were readily identified as HCs by IF. Detection sensitivity of spiked cancer cells in donor blood was similar for both methods.

Conclusions

There is a high frequency of CK+ events in blood and marrow, and it is important to note that this is observed both in patients with and those without cancer. IF with multiple HC markers allows straightforward discrimination between CK+ cells of hematopoietic and nonhematopoietic origin.

Keywords

ImmunocytochemistryBone marrowCytokeratinImmunofluorescence

Copyright information

© The Society of Surgical Oncology, Inc. 2005