The AdnaTest uses a two-step process (Select and Detect) to generate results within 5 hours (see “AdnaTest CTC-Select: Immunomagnetic cell selection with multiple tumor associated antibodies.
” and “AdnaTest LungCancerDetect for analysis of various cancer-associated tumor markers.
”). AdnaTest CTC-Select
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), and subsequently lysed. (see “AdnaTest Select: Immunomagnetic cell selection with multiple tumor-associated antibodies labeled to magnetic beads
”). The cell lysate is then used in AdnaTest LungCancerDetect
, step 2 of the procedure, to isolate mRNA.
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 (See "AdnaTest LungCancerDetect for analysis of various cancer-associated tumor markers.
Multiplex PCR provides tumor-associated gene expression profiles for a variety of relevant tumor markers to ensure that the selected cells are cancer cells (see “AdnaTest LungCancerDetect results of samples using an Agilent 2100 Bioanalyzer
”). The kit contains one primer mix to amplify cDNA target fragments of Twist, PI3Kα, Akt2 and a housekeeping gene as control (actin) (see "AdnaTest LungCancer specificity and recovery
."). The PrimerMix LungDetect amplifies ALDH1 and EpCam (GA733-2) (below). The primers generate fragments of the following sizes:
Primer Mix EM-2 Detect
|Target gene ||Fragment size |
|PI3Kα ||551 bp |
|Akt-2 ||309 bp |
|TWIST1 ||201 bp |
|Actin ||120 bp (internal PCR control) |
Primer Mix LungDetect
|Target Gene ||PCR fragment size |
|EpCam (GA733-2) ||395 bp |
|ALDH1 ||161 bp |
Importantly, the AdnaTest is an open platform that allows the profiling of additional custom selected mRNA targets for comprehensive biomarker discovery.