UNASSIGNED: Circulating @entity5 cells (CTCs) are shed from a solid @entity5 into the bloodstream and can seed new metastases. CTCs hold promise for @entity5 diagnosis and prognosis and to increase our understanding of the metastatic process. However, their low numbers in blood and varied phenotypic characteristics make their detection and isolation difficult. One source of heterogeneity among CTCs is molecular: When they leave the @entity5 , these cells must undergo a molecular transition, which increases their mobility and chance of survival in the blood. During this molecular transition, the cells lose some of their epithelial character, which is manifested by the expression of the @entity3058 known as @entity5960 ( @entity5960 ). Some @entity5 shed CTCs that express high levels of @entity5960 ; others release cells that have a low level of the antigen. Nevertheless, many CTC isolation techniques rely on the detection of @entity5960 to discriminate CTCs from other cells in the blood. We previously reported a high-throughput immunofluorescence-based technology that targets @entity5960 to rank aliquots of blood for the presence or absence of a CTC. This technology, termed ensemble decision aliquot ranking (eDAR), recovered spiked-in @entity5 cells (taken from a model @entity5960 (high) cell line) from blood at an efficiency of 95%. In this paper, we evaluated eDAR for recovery of cells that have low @entity5960 expression and developed an immunofluorescence labeling strategy that significantly enhances the method's performance. Specifically, we used a cocktail of primary antibodies for both epithelial and mesenchymal antigens as well as a dye-linked secondary antibody. The cocktail allowed us to reliably detect a model @entity5960 (low) cell line for triple negative @entity0 , MDA-MB-231, with a recovery efficiency of 86%. Most significantly, we observed an average of 6-fold increase in the number of CTCs isolated from blood samples from @entity0 @entity1 . These findings underscore the importance of benchmarking CTC technologies with model cell lines that express both high and low levels of @entity5960 .

Improved Detection by Ensemble-Decision Aliquot Ranking of Circulating XXXX Cells with Low Numbers of a Targeted Surface Antigen.

@entity1 :: ('9606', 'Species') :: ['patients'] @entity5960 :: ('4072', 'Gene') :: ['epithelial cell adhesion molecule', 'EpCAM'] @entity0 :: ('MESH:D001943', 'Disease') :: ['breast cancer'] @entity5 :: ('MESH:D009369', 'Disease') :: ['tumor', 'cancer', 'primary tumor', 'tumors'] @entity3058 :: ('963', 'Gene') :: ['cell surface antigen']
@entity5 :: (MESH:D009369,Disease) :: ['Tumor']
