Comparison of assay methods for detection of circulating tumor cells in metastatic breast cancer: AdnaGen AdnaTest BreastCancer Select/Detect™ versus Veridex CellSearch™ system

Andreopoulou, Eleni; Yang, Liangpeng; Rangel, Kelly; Reuben, James M.; Hsu, Limin; Krishnamurthy, Savitri; Valero, Vicente; Fritsche, Herbert A.; Cristofanilli, Massimo

doi:10.1002/ijc.26111

Cited by 195 publications

(132 citation statements)

References 33 publications

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“…Recently, Markou et al described a liquid bead array hybridization assay in which mRNA isolated from immunomagnetically enriched CTCs are subjected to multiplex PCR for simultaneous measurements of the expression of 6 genes in CTCs (20 ). A commercially available RNA-based CTC assay, the AdnaTest™ (AdnaGen), uses nonquantitative RT-PCR to identify cells that express the putative transcripts of tumor-specific genes after immunomagnetic capture of MUC1 ϩ /HER2 ϩ / EpCAM ϩ cells (21 ). The assay's limitations, which derive from the fact that MUC1 is also expressed on activated T lymphocytes, should be considered (22 ).…”

Section: Mrna-based Strategiessupporting

confidence: 44%

Circulating Tumor Cells: Liquid Biopsy of Cancer

2013

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BACKGROUND:The detection and molecular characterization of circulating tumor cells (CTCs) are one of the most active areas of translational cancer research, with Ͼ400 clinical studies having included CTCs as a biomarker. The aims of research on CTCs include (a) estimation of the risk for metastatic relapse or metastatic progression (prognostic information), (b) stratification and real-time monitoring of therapies, (c) identification of therapeutic targets and resistance mechanisms, and (d) understanding metastasis development in cancer patients.CONTENT: This review focuses on the technologies used for the enrichment and detection of CTCs. We outline and discuss the current technologies that are based on exploiting the physical and biological properties of CTCs. A number of innovative technologies to improve methods for CTC detection have recently been developed, including CTC microchips, filtration devices, quantitative reverse-transcription PCR assays, and automated microscopy systems. Molecularcharacterization studies have indicated, however, that CTCs are very heterogeneous, a finding that underscores the need for multiplex approaches to capture all of the relevant CTC subsets. We therefore emphasize the current challenges of increasing the yield and detection of CTCs that have undergone an epithelialmesenchymal transition. Increasing assay analytical sensitivity may lead, however, to a decrease in analytical specificity (e.g., through the detection of circulating normal epithelial cells).

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Section: Mrna-based Strategiessupporting

confidence: 44%

Circulating Tumor Cells: Liquid Biopsy of Cancer

2013

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“…Detection of CTCs in blood is a noninvasive method of sampling and studying tumour cells compared to classic biopsy. Most CTCs enrichment methods are based on the use of EpCAM-specific antibodies and subsequent detection using anti-cytokeratin [13,14,29,30]. EpCAM is a surface marker found only on carcinoma cells.…”

Section: Discussionsupporting

confidence: 42%

Analysis of the specificity and selectivity of anti-EpCAM antibodies in breast cancer cell lines

Sterzyńska

Kempisty²,

Zawierucha³

et al. 2012

Folia Histochem Cytobiol.

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Abstract:The epithelial cell adhesion molecule (EpCAM) is a membrane glycoprotein that is expressed in most normal human epithelia and overexpressed in most carcinomas. This molecule is responsible for cell-to-cell adhesion and additionally participates in signalling, cell migration, proliferation and differentiation. Therefore, EpCAM has been the target of immunotherapy in clinical trials of several solid tumours. It has been found to play an important role in the detection and isolation of circulating tumour cells (CTCs). The aim of this study was to investigate and compare the specificity and selectivity of different anti-EpCAM antibodies in order to assess their usefulness for CTCs binding. All experiments were performed in six different types of breast cancer cell lines (MCF-7, SkBr-3, T47D, CAMA-1, MDAMB-231, and BT-20) and using three different anti-EpCAM antibodies (EBA-1, AUA-1, and 9C4). Immunofluorescence and Real-Time PCR techniques were applied to analyse the protein and gene expression levels. The experiments revealed that the investigated antibodies differed significantly regarding the specificity of EpCAM antigen binding. The most significant role in targeting CTCs was played by the EBA-1 and 9C4 anti-EpCAM antibodies. They revealed the strongest immunofluorescent signal among other applied antibodies and/or were specific for all examined breast cancer cell lines. The strength and specificity of reaction was dependent not only on the type of antibody, but also on the type of breast cancer cell line. We noted that the diverse sensitivities of reactions depended on the type of applied antibody. We therefore recommend the simultaneous application of different anti-EpCAM antibodies. An appropriate choice of anti-EpCAM antibodies and an evaluation of EpCAM expression in breast cancer appear to be crucial, especially as this antigen is being proposed as a marker for the detection of circulating tumour cells.

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“…AdnaTest used RT-PCR to detect gene transcripts of tumor markers (GA733-2, MUC-1, and HER2). The results indicated that AdnaTest has a sensitivity equivalent to that of the CellSearch system (Andreopoulou et al, 2012). Kim et al used a telomerase-specific replication-selective adenovirus to detect CTCs based on the principle that the adenovirus can replicate only in telomeraseexpressing cells and emit fluorescence in transfected cells.…”

Section: Liquid Biopsies In Tumour Diagnosis and Treatmentsupporting

confidence: 40%

Liquid biopsies: tumour diagnosis and treatment monitoring

Pham

2016

Biomed Res Ther

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Abstract-Cancer is a disease with high evolutionary, i.e., malignant, characteristics that change under selective pressure from therapy. Characterization based on molecular or primary tumor properties or clinicopathological staging does not fully reflect the state of cancer, especially when cancer cells metastasize. This is the major reason for failure of cancer treatment. Currently, there is an urgent need for new approaches that allow more effective, but less invasive, monitoring of cancer status, thereby improving the efficacy of treatments. With recent technological advances, "liquid biopsies," the isolation of intact cells or analysis of components that are secreted from cells, such as nucleic acids or exosomes, could be implemented easily. This approach would facilitate real-time monitoring and accurate measurement of critical biomarkers. In this review, we summarize the recent progress in the identification of circulating tumor cells using new high-resolution approaches and discuss new circulating tumor nucleic acid-and exosome-based approaches. The information obtained through liquid biopsies could be used to gain a better understanding of cancer cell invasiveness and metastatic competence, which would then benefit translational applications such as personalized medicine.

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Comparison of assay methods for detection of circulating tumor cells in metastatic breast cancer: AdnaGen AdnaTest BreastCancer Select/Detect™ versus Veridex CellSearch™ system

Cited by 195 publications

References 33 publications

Circulating Tumor Cells: Liquid Biopsy of Cancer

Circulating Tumor Cells: Liquid Biopsy of Cancer

Analysis of the specificity and selectivity of anti-EpCAM antibodies in breast cancer cell lines

Liquid biopsies: tumour diagnosis and treatment monitoring

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