Qiagen, 395022, AdnaTest ColonCancerSelect

For isolation of CTCs and the subsequent extraction of mRNA from human whole blood for 12 preparations

Features

- Efficient isolation and detection of CTCs from whole blood
- Molecular characterization of CTCs
- High specificity and sensitivity
- Processing of multiple samples in parallel

Procedure

Target gene: PCR Fragment Size (bp)
GA733-2: 383
CEA: 231
EGFR: 163
Actin (internal PCR control): 120

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:

Importantly, the AdnaTest is an open platform that allows the profiling of additional custom selected mRNA targets for comphrehensive biomarker discovery.

Applications

- Isolation of CTCs from whole human blood
- Molecular characterization of CTCs
- Biomarker discovery
- mRNA profiling
- Liquid biopsy