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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.