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Features
- Efficient isolation and detection of CTCs from whole blood
- Molecular characterization of CTCs
- High specificity and sensitivity
- Processing of multiple samples in parallel
Product Details
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.
Performance
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.
Principle
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).
Procedure
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:
| 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 comphrehensive biomarker discovery.
Applications
- Isolation of CTCs from whole human blood
- Molecular characterization of CTCs
- Biomarker discovery
- mRNA profiling
- Liquid biopsy
Supporting data and figures
AdnaTest BreastCancerDetect results
The first lane shows the DNA size standard
(DNA-Ladder). Sample 1 is positive for GA733-2, Muc-1 and Her-2. Samples 3, 4 and 5 are
positive for GA733-2. Muc-1 in sample 2 is negative. The only band detected in sample 6 is for
GA733-2. Actin is detected in samples 1 to 6. The PCR negative (C-) and positive control (C+)
are shown in the last two lanes (Agilent 2100 Bioanalyzer).
Resources
MSDS (1)
Safety Data Sheets (1)