Qiagen, 396132, AdnaTest ProstateCancerPanel AR-V7

For 12 enrichments of tumor cells from whole blood and subsequent detection of prostate cancer-associated gene expression including AR-V7 expression.

Features

- Efficient isolation and detection of CTCs from whole blood
- Molecular characterization of CTCs
- High specificity and sensitivity

Procedure

Recovery [%]: n
2 tumor cells spiked into 5 ml blood: 38/40 (95%)

PSA: PSMA
Cut-off: 35.00
Positives after cut-off: 0
Specificity: 100%

PSA: PSMA
Slope over the range: -3.55
Efficiency: -1+10(-1/slope): 91%
Amplification factor: 10^(1/-slope): 1.91

Recovery

The AdnaTest ProstateCancerSelect is used for the enrichment of CTCs from whole blood in prostate cancer research. Subsequently, the AdnaTest ProstateCancerPanel AR-V7 is used for molecular characterization by analyzing gene expression of AR-V7 and further prostate cancer associated genes. In spiking experiments, 2 tumor cells in 5 ml of whole blood are detected at a recovery rate of 95%. >

Two cancer cells (Prostate cancer cell line LnCap) were spiked into blood samples (5 ml) from healthy donors (n=40) to determine the recovery rates achieved with the AdnaTest ProstateCancerPanel AR-V7 as summarized in above. >

Specificity

Fourteen male healthy donors were analyzed with the AdnaTest ProstateCancerPanel AR-V7 to determine the rate of false positives/specificity at the given cut offs. >

The AdnaTest ProstateCancerPanel AR-V7 could demonstrate in this performance evaluation a specificity of 100% for all tumor associated genes (PSA, PSMA, AR-V7) except ARwt where it reached 93% (details in table above). Also, GAPDH reached 100% specificity and CD45 79%, respectively, whereas these 2 genes are not used for CTC characterization as such, hence a specificity below 90% for those two target genes is acceptable. >

qRT-PCR linearity and efficiency

Perfect linearity and efficiency of the qRT-PCR reactions for each of the genes contained in the AdnaTest ProstateCancerPanel AR-V7 was proven in a serial dilution series of standards (See “ AR-V7 Panel gene dilutions ”). The positive control has been used as template for standard sample generation. ​ The resulting Ct values for this dilution series were clearly linear, correlated to the copy numbers for each gene at each dilution step (See “ AR-V7 Panel gene dilutions ”) getting down to ~500 copies. The qRT-PCR efficiency of all the tumor cell related genes was higher than 90% and the corresponding amplification factors ranged above 1.9 which all in all can be taken as a prove for very effective and sensitive qRT-PCR design (see the table below).>

Field test results

The AdnaTest ProstateCancerPanel AR-V7 workflow was tested in 12 castration-resistant prostate cancer (CRPC) samples. See " Gene profile in clinical samples .">

Applications

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