therascreen PITX2 RGQ PCR Kit
For clinically reliable determination of the PITX2 methylation ratio in lymph node-positive, estrogen receptor-positive and HER2-negative high-risk breast cancer patient tissue samples
For clinically reliable determination of the PITX2 methylation ratio in lymph node-positive, estrogen receptor-positive and HER2-negative high-risk breast cancer patient tissue samples
Cat. No. / ID: 9024203
The therascreen PITX2 RGQ PCR Kit is a CE-IVD kit for determination of the percent methylation ratio (PMR) in promoter 2 of the pituitary homeobox transcription factor 2 (PITX2) gene using methylation-specific real-time PCR on the Rotor-Gene Q 5-plex HRM instrument. The kit provides reagents optimized for reliable and sensitive determination of the PITX2 methylation status, and allows for a clinically validated determination of the PMR cut-off value of 12, which is predictive of patient response to anthracycline-based chemotherapy with or without endocrine therapy when used to test formalin-fixed, paraffin-embedded (FFPE) tissue samples from lymph node-positive, estrogen receptor-positive and HER2-negative high-risk breast cancer patients. Want to try this solution for the first time? Request a quote for a trial kit.
The therascreen PITX2 RGQ PCR Kit exploits the real-time PCR oligonucleotide hydrolysis principle of TaqMan probes in combination with methylation-unspecific primers. Distinction between methylated and unmethylated status of the amplified bisulfite-converted target sequence in the PITX2 promoter 2 is achieved by using the combination of a FAM-labeled probe hybridizing specifically to the methylated bisulfite-converted DNA sequence and a HEX-labeled probe specific for the unmethylated bisulfite-converted DNA sequence. The resulting FAM- and HEX-specific Ct values are then used for the automated calculation of the PMR.
The first step is manual DNA extraction using the QIAamp DSP DNA FFPE Tissue Kit, followed by quantification of genomic DNA and then bisulfite conversion with EpiTect Fast DNA Bisulfite Kit. Bisulfite-converted DNA is then amplified by real-time PCR, with all reagents provided in the therascreen PITX2 RGQ PCR Kit and the methylation status of the PITX2 promoter 2 is determined using the Rotor-Gene Q MDx 5plex HRM instrument. Finally, the Rotor-Gene AssayManager v2.1 software is used to automatically determine the run/sample validity and calculate the PMR.
The therascreen PITX2 RGQ PCR Kit enables the determination of the percent methylation ratio (PMR) in promoter 2 of the PITX2 gene. This PMR value of PITX2 promotor 2 is predictive of the response to anthracycline-based chemotherapy regimes in lymph node-positive, estrogen receptor-positive and HER2-negative high-risk breast cancer patients. Applying PITX2 methylation as a predictive biomarker can help clinicians to select the appropriate chemotherapy regimen.
The therascreen PITX2 RGQ PCR Kit is compliant with EU IVD Directive 98/79/EC, enabling the determination of the percent methylation ratio of PITX2 promoter 2 for in vitro diagnostics.