Description
The EZ2 AdnaTest CTC Select Kit provides immunomagnetic enrichment of circulating tumor cells from human whole blood on the EZ2 Connect instrument. Integrated mRNA isolation allows molecular characterization of CTCs using RT-PCR, real-time RT-PCR, or cDNA generation, including the AdnaTest procedure for molecular characterization of CTCs.
EZ2 AdnaTest CTC Select Kit allows the automated enrichment of CTCs from human whole blood based on the AdnaTest principle using the EZ2 Connect instrument. The EZ2 AdnaTest CTC Select Kit is a highly specific immunomagnetic cell-selection system for enriching circulating tumor cells (see Table "Performance comparable to manual protocol – EZ2 AdnaTest CTC Select specificity and recovery") from peripheral blood including integrated mRNA isolation. The EZ2 Connect automates the CTC enrichment and subsequent mRNA isolation using prefilled cartridges. The benchtop instrument automatically pierces the sealed cartridges, and automates pipetting of beads and buffers. Its intuitive touchscreen interface makes it easy to use, even for new lab workers. And with its remote connectivity feature, users can monitor runs and the instrument can be managed even from outside the lab (see figure "Easy to use – enrich circulating tumor cells for molecular analyses ”). Performance comparable to manual protocol – EZ2 AdnaTest CTC Select specificity and recovery Method Manual Automated AdnaTest CTC Select and BreastCancer Add-on EZ2 AdnaTest CTC Select and BreastCancer Add-on Genes Her2 (265 bp) Muc1 (299 bp) Ga733-2 (395 bp) Her2 (265 bp) Muc1 (299 bp) Ga733-2 (395 bp) Non-spiked healthy donor samples 0/6 0/6 Specificity 100% 100% Spiked healthy donor samples 6/6 6/6 Recovery 100% 100% Blood samples (5 ml) from six healthy donors (n=6) were analyzed directly or spiked with five cancer cells (breast cancer cell line MCF-7). Samples were prepared using the manual AdnaTest CTC Select Kit or the automated version of the AdnaTest, the EZ2 AdnaTest CTC Select Kit on the EZ2 Connect. Sensitivity and specificity were determined using the AdnaTest PrimerMix BreastDetect which allows amplification of three tumor-associated antigens and one control gene. The high-quality mRNA purified has proven identical specificity and sensitivity in downstream applications to mRNA isolated by the manual protocol, and can therefore be used in any application, such as RT-PCR, real-time RT-PCR, or cDNA generation, including the AdnaTest procedure for molecular characterization of CTCs. In combination with EMT-2 Add-on CancerDetect products, the resulting mRNA/cDNA can therefore be used to generate results for the following cancer entities: Breast cancer (see figure "Breast cancer-associated gene expression") Prostate cancer (see figure "AdnaTest prostate cancer-associated gene expression "and "Gene profile in clinical samples") Ovarian cancer (see figure "AdnaTest ovarian cancer results") Lung cancer (see figure "Lung cancer- and EMT-2-associated gene expression") Colon cancer (see figure "Colon cancer-associated gene expression") Tumor stem cells (see figure "Tumor stem cell- and EMT-2-associated gene expression")
EZ2 AdnaTest CTC Select provides prefilled cartridges containing all buffers for the enrichment and lysis of CTCs. A fresh blood sample is incubated with CTC Select beads before loading the sample into the EZ2 Connect instrument. The instrument automatically pierces the prefilled cartridges and enriches the tumor cells using in-tip magnetic bead separation. 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 the EZ2 Connect and lysed. The cartridges also contain oligo (dT) 25-coated beads for isolation of mRNA from the lysate of pre-enriched tumor cells (see figure "During the run"). Reverse transcription results in cDNA that can be used as template for tumor cell detection and characterization by PCR. Importantly, the AdnaTest is an open platform that allows the profiling of additional custom-selected mRNA targets for comprehensive biomarker discovery.
Fresh blood samples are incubated with CTC Select beads before being loaded into the EZ2 Connect instrument. Oligo dT beads and an elution tube are added to the prefilled EZ2 AdnaTest CTC Select cartridges before loading together with the preincubated samples into the EZ2 Connect instrument. Instrument setup is finalized by adding the large volume rack (cat. no. 9027011) and adding tips and tip holders (provided separately, cat. no. 994900). The EZ2 AdnaTest CTC Select protocol is selected and the instrument started (see figure: Getting started).
