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Annals of Surgical Oncology
Published in: Annals of Surgical Oncology 9/2005

01-09-2005

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

Authors: David N. Krag, MD, FACS, Roberto Kusminsky, MD, Edward Manna, BS, Abiy Ambaye, MD, Donald L. Weaver, MD, Seth P. Harlow, MD, Michael Covelli, MD, Mary A. Stanley, MD, Laurence McCahill, MD, Frank Ittleman, MD, Bruce Leavitt, MD, Martin Krag, MD

Published in: Annals of Surgical Oncology | Issue 9/2005

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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.
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Metadata
Title
The Detection of Isolated Tumor Cells in Bone Marrow Comparing Bright-Field Immunocytochemistry and Multicolor Immunofluorescence
Authors
David N. Krag, MD, FACS
Roberto Kusminsky, MD
Edward Manna, BS
Abiy Ambaye, MD
Donald L. Weaver, MD
Seth P. Harlow, MD
Michael Covelli, MD
Mary A. Stanley, MD
Laurence McCahill, MD
Frank Ittleman, MD
Bruce Leavitt, MD
Martin Krag, MD
Publication date
01-09-2005
Publisher
Springer-Verlag
Published in
Annals of Surgical Oncology / Issue 9/2005
Print ISSN: 1068-9265
Electronic ISSN: 1534-4681
DOI
https://doi.org/10.1245/ASO.2005.12.004

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