Abstract
Metastasis describes the process of migration of a frequently clinically occult circulating tumor cell (CTC) from the primary lesion to a new location and the subsequent formation of an overt growth. We and others have shown that the detection and quantitation of these cells has significant prognostic value, however there still remains no consensus as to the optimal methods to achieve this. The work described herein therefore considered various techniques, from storage and sample processing to data acquisition and analysis, to find an optimal combination of methods for an effective and practical platform for the detection of CTCs in peripheral blood. A dual-antigen epithelial cell enrichment procedure followed by a multi-marker QPCR analysis demonstrated the highest sensitivity and specificity, with the ability to detect as few as 10 tumor cells from a background of 10(6) peripheral blood mononuclear cells. Using these techniques in conjunction with a quadratic linear discriminant analysis (QDA) resulted in a platform able to generate this data and then combine it a single score for each patient, in which positivity reflected tumor cell presence, and negativity represented tumor cell absence. This assay was able to correctly de...Continue Reading
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