Description
AdnaTest ColonCancer allows molecular characterization of CTCs by combining AdnaTest ColonCancerSelect with AdnaTest ColonCancerDetect using the Combination of Combinations Principle (COCP). AdnaTest ColonCancerSelect is a highly specific immunomagnetic cell-selection system for enriching circulating tumor cells from peripheral blood. AdnaTest ColonCancerDetect allows sensitive analysis of colon cancer-associated gene expression in immunomagnetically enriched tumor cells by reverse transcription and PCR.
AdnaTest ColonCancer 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 colon 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, CEA and EGFR.
The AdnaTest uses a two-step process (Select and Detect) to generate results within 5 hours (see figures “ AdnaTest workflow ”). AdnaTest ColonCancerSelect 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 ColonCancerDetect 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 ColonDetect allows amplification of 3 tumor-associated genes and 1 control gene (see table below). The primers generate fragments of the following sizes: PrimerMix ColonDetect Target gene PCR Fragment Size (bp) GA733-2 383 CEA 231 EGFR 163 Actin (internal PCR control) 120 Importantly, the AdnaTest is an open platform that allows the profiling of additional custom selected mRNA targets for comphrehensive biomarker discovery.
