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Comparing Sequential Steps For Detection Of Circulating Tumor Cells: More Specific Or Just Less Sensitive?

Submitted on:08-Feb-2011, 08:08:57 AM GMT Published on: 08-Feb-2011, 09:38:27 PM GMT

Article URL: http://www.webmedcentral.com/article_view/1490

The corresponding author Dr. Pachmann holds a patent 7615358 protecting the MAINTRAC method outside of Europe and is partner in the SIMFO GmbH a company dedicated at developing new tests but which does not sell tests to patients. Tests are performed for physicians and patients on request as medically indicated and supervised tests by the transfusion-medical laboratory Dr. Ulrich Pachmann.

Comparing Sequential Steps For Detection Of Circulating Tumor Cells: More Specific Or Just Less Sensitive?

Author(s):Dr. Ulrich A Pachmann, Mrs. Katya Hekimian, Mrs. Stephanie Carl, Mrs. Nadine Ruediger, Mrs.
Carola Rabenstein, Prof. Katharina Pachmann
Corresponding Author:
Dr. Ulrich A Pachmann,
Head of the Institute, Transfusion Center Bayreuth, Kurpromenade 2, 95448 - Germany
Submitting Author:
Dr. Ulrich A Pachmann,
Head of the Institute, Transfusion Center Bayreuth, Kurpromenade 2, 95448 - Germany
Article ID: WMC001490
Article Type: Research Protocol
Submitted on:08-Feb-2011, 08:08:57 AM GMT Published on: 08-Feb-2011, 09:38:27 PM GMT
Article URL: http://www.webmedcentral.com/article_view/1490
Subject Categories:CANCER
Keywords:Circulating epithelial tumor cells, Detection methods, Sensitivity,
How to cite the article:Pachmann U A, Hekimian K , Carl S , Ruediger N , Rabenstein C , Pachmann K .
Comparing Sequential Steps For Detection Of Circulating Tumor Cells: More Specific Or Just Less Sensitive? .
WebmedCentral CANCER 2011;2(2):WMC001490
Source(s) of Funding:
No current external funding sources for this study
Competing Interests:
The corresponding author Dr. Pachmann holds a patent 7615358 protecting the MAINTRAC method outside of
Europe and is partner in the SIMFO GmbH a company dedicated at developing new tests but which does not sell
tests to patients. Tests are performed for physicians and supervised tests by the
transfusion-medical laboratory Dr. Ulrich Pachmann.


Abstract

Background: We compared two surface epithelial antigen (EpCAM)-based approaches for the detection of breast cancer cells present in the circulation.

Methods: Blood from 20 breast cancer patients was drawn into standard blood collection tubes (SBCT) and of 7 of these additionally into CellSave® tubes. After erythrocyte lysis of the samples from both systems, cells were stained with FITC-anti-EpCAM and propidium iodide, quantified with an automated microscope and intact cells counted.

Results: EpCAM-positive events from 1ml of blood ranged between 2051 and 28875/ml and from SBCT (MAINTRAC® approach) as compared to between 97 and 2343/ml from the CellSave® tubes, indicating a more than 10-fold reduction in EpCAM accessibility by the preservative. Duplicate cell preparations showed a high correlation of R2=0.89 (MAINTRAC®) from SBCT vs. a moderate correlation of R2=0.81 from CellSave® tubes, but a good correlation (R2=0.91) between the events detected from both systems.

Between 1/2 and 1/5 of the positive events were viable cells in the MAINTRAC® approach with unequivocal morphology, and a good (R2=0.89) correlation to total events; by contrast, 1/10 to less than 1/100 of the events in the CellSave® tubes were perhaps cells with equivocal morphology no correlation to total events most positive events being non-recognizable cells. Still 30 to 100-fold more cells were recovered than with the CellTrack® Analyzer.

Conclusions: The approach without fixative detects considerably more EpCAM-positive events with good cell morphology as compared to the CellSave® fixation where cell morphology is poor. Magnetic bead enrichment further reduces the number of retrieved cells.

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