Description
AdnaTest BreastCancer allows molecular characterization of CTCs by combining AdnaTest BreastCancerSelect with AdnaTest BreastCancerDetect using the Combination of Combinations Principle (COCP). AdnaTest BreastCancerSelect is a highly specific immunomagnetic cell-selection system for enriching circulating tumor cells from peripheral blood. AdnaTest BreastCancerDetect allows sensitive analysis of breast cancer-associated gene expression in immunomagnetically enriched tumor cells by reverse transcription and PCR.
AdnaTest BreastCancer is a highly specific immunomagnetic cell selection system for enriching circulating tumor cells from peripheral blood and includes a highly sensitive RT-PCR for detecting breast cancer-specific mRNA markers.
AdnaTest technology uses an optimized combination of antibodies for cell selection in 5 ml whole blood and a combination of tumor-associated markers for the detection of tumor cells: EpCAM, CA15-3 (Muc1), presence of Her2 receptors (optional detection of estrogen and progesterone receptors by AdnaTest ER / PR Detect).
The AdnaTest uses a two-step process (Select and Detect) to generate results within 5 hours (see figures “ AdnaTest workflow ”). AdnaTest BreastCancerSelect enables the immunomagnetic enrichment of tumor cells from whole blood via epithelial- and tumor-associated antigens. Antibodies against epithelial- and tumor-associated antigens are conjugated to magnetic beads for labeling of tumor cells. Labeled cells are extracted by a magnetic particle concentrator (AdnaMag-L and AdnaMag-S), lysed and mRNA is then purified from these lysates (see figures “ AdnaTest Select – Immunomagnetic cell selection with multiple tumor-associated antibodies labeled to magnetic beads ”). In the second step, the isolated mRNA is transcribed into cDNA and can be amplified using multiplex PCR (see figures “ AdnaTest Detect – Multiplex PCR of various cancer-associated tumor markers ”). AdnaTest BreastCancerDetect contains Oligo (dT) 25-coated beads for the isolation of mRNA from the lysate of pre-enriched tumor cells. Reverse transcription results in cDNA that can subsequently be used as template for tumor cell detection and characterization by multiplex PCR. Multiplex PCR provides tumor-associated gene expression profiles for a variety of relevant tumor markers to ensure that the selected cells are cancer cells. The PrimerMix BreastDetect allows amplification of 3 tumor-associated genes and 1 control gene (see table below). The primers generate fragments of the following sizes: PrimerMix BreastDetect Target gene PCR fragment size (bp) GA733-2 395 Muc-1 299 Her-2 265 Actin (internal PCR control) 120 Importantly, the AdnaTest is an open platform that allows the profiling of additional custom selected mRNA targets for comprehensive biomarker discovery.
