Qiagen, 399932, AdnaTube

12 sample tubes containing EDTA. Use only with anticoagulated blood collected in ACDA blood collection tubes from BD (Becton Dickinson, 8.5 ml)

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)
PSMA: 449
PSA: 357
EGFR: 163
Actin (internal PCR control): 120

Target gene: PCR fragment size (bp)
AR: 440

The AdnaTest uses a two-step process (Select and Detect) to generate results within 5 hours (see figures “ AdnaTest workflow ”). AdnaTest ProstateCancerSelect 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 ProstateCancerDetect 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 ProstateDetect allows the amplification of three tumor associated antigens and one control gene. With the PrimerMix AR-Detect the androgen receptor (AR) is amplified. 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