detection of epcam-negative and cytokeratin-negative circulating tumor cells in peripheral blood检测epcam-negative和cytokeratin-negative外周血循环肿瘤细胞.pdfVIP

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detection of epcam-negative and cytokeratin-negative circulating tumor cells in peripheral blood检测epcam-negative和cytokeratin-negative外周血循环肿瘤细胞.pdf

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detection of epcam-negative and cytokeratin-negative circulating tumor cells in peripheral blood检测epcam-negative和cytokeratin-negative外周血循环肿瘤细胞

Hindawi Publishing Corporation Journal of Oncology Volume 2011, Article ID 252361, 10 pages doi:10.1155/2011/252361 Research Article Detection of EpCAM-Negative and Cytokeratin-Negative Circulating Tumor Cells in Peripheral Blood Stephen D. Mikolajczyk,1 Lisa S. Millar,1 Pavel Tsinberg,1 Stephen M. Coutts,1 Maryam Zomorrodi,1 Tam Pham,2 Farideh Z. Bischoff,2 and Tony J. Pircher1 1 Research and Development, Biocept Inc., 5810 Nancy Ridge Drive, Suite 150, San Diego, CA 92121, USA 2 Translational and Clinical Development, Biocept Inc., 5810 Nancy Ridge Drive, Suite 150, San Diego, CA 92121, USA Correspondence should be addressed to Stephen D. Mikolajczyk, smikolajczyk@ Received 4 January 2011; Accepted 14 February 2011 Academic Editor: Michael Carducci Copyright © 2011 Stephen D. Mikolajczyk et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Enrichment of rare circulating tumor cells (CTCs) in blood is typically achieved using antibodies to epithelial cell adhesion molecule (EpCAM), with detection using cytokeratin (CK) antibodies. However, EpCAM and CK are not expressed in some tumors and can be downregulated during epithelial-to-mesenchymal transition. A micro-ﬂuidic system, not limited to EpCAM or CK, was developed to use multiple antibodies for capture followed by detection using CEE-Enhanced (CE), a novel in situ staining method that ﬂuorescently labels the capture antibodies bound to CTCs. Higher recovery of CTCs was demonstrated using antibody mixtures compared to anti-EpCAM. In addition, CK-positive breast cancer cells were found in 15 of 24 samples (63%;