SensiScreen® and PlentiPlex™ MYD88 are real-time PCR-based assays for somatic mutation detection in cancer patients. SensiScreen® and PlentiPlex™ MYD88 combine high sensitivity with ease-of-use and are designed to work on standard real-time PCR equipment. Sensitive detection of somatic mutations by SensiScreen® and PlentiPlex™ MYD88 is achieved with PentaBase’s novel and selective INA® technologies including the use of BaseBlockers™ that suppress false positive signals from wild type templates.
From sample to answer in less than three hours
SensiScreen® and PlentiPlex™ MYD88 assays are supplied as either “Ready-to-use” or as “Dispense Ready” mixes. Ready-to-use assays are pre-dispensed in PCR strips and require only addition of gDNA for somatic mutation testing. Dispense-ready mixes include oligonucleotides and master mix in separate tubes ready to be dispensed and added gDNA. The technologies are applicable on standard real-time equipment using standard procedures and results can be ready in less than three hours from receiving a sample (including automated DNA extraction).
Ultra-sensitive detection of somatic mutations by SensiScreen® and PlentiPlex™ MYD88 is possible with PentaBase’s novel and selective technologies, comprising:
1. HydrolEasy™ probes with significantly improved signal-to-noise ratio and higher specificity compared to conventional hydrolysis probes.
2. Suprimer™ DNA primers modified with pentabases for specific and sensitive amplification.
3. BaseBlockers™ suppressing false positive signals from wild type templates and ensuring high specificity. and robustness of the assays.
• Solid and liquid sample analysis: SensiScreen® assays are supplied in both an FFPE version for analysis of solid biopsies and a liquid version for analysis of liquid biopsies containing small amounts of DNA.
• High Sensitivity: PlentiPlex™ MYD88 and most SensiScreen® FFPE assays have a limit of detection of mutated DNA below 1% in a wild type background. SensiScreen® Liquid assays detect down to one copy of mutated DNA.
• Easy and fast: Easy to setup (hands-on-time <2 min/sample) and it takes less than three hours from sample to result including DNA extraction .
• State of the art: Based on PentaBase’s proprietary INA® technology.
• Proven: CE-IVD marked assays are clinically validated in Switzerland and Denmark, and are certified in accordance to the EU guidelines 98/79/EC Medical equipment for in vitro diagnostics.
• Open qPCR platform designs: Designed to work on standard real-time PCR instruments and validated on commonly used platforms.
SensiScreen® Liquid assays are designed for early detection and monitoring of somatic mutations in liquid biopsies using standard equipment. With SensiScreen® Liquid, it is possible to monitor changing levels of cancer mutations and detect newly developed mutations in circulating cell-free DNA (ccfDNA) shed into the blood or other liquid samples. In this way, SensiScreen® Liquid can support the selection of the most appropriate personalized treatment in a timely manner. SensiScreen® Liquid assays are able to detect down to a single copy of mutated DNA in a background of wild type DNA.
Detecting down to one copy
The figures below illustrates a SensiScreen® Liquid assay detecting one copy of mutant DNA. 12 identical of a patient DNA Heterozygous for an EGFR Exon 19 Deletion divided to approximately 3 copies per. well were carried out.
Detection in solid biopsies
SensiScreen® FFPE assays are designed for detection of somatic mutations from genomic DNA in solid biopsies (fresh, frozen or FFPE tissue) or other samples with high amounts of DNA (load up to 50 ng).
SensiScreen® FFPE assays are able to robustly detect 0.25-1 % mutation in a 50 ng wild type background.
Rotor-Gene 6000 PCR amplification plots of SensiScreen® KRAS G12V simplex and multiplex assays using serial dilutions of mutated DNA in a wild type background. 50 ng (about 16.000 copies) of human genomic DNA was added to each reaction. The threshold was set at 10% of the average fluorescence signal of the reference assay at cycle 45. Legend describes the fraction of mutated G12V cell line DNA in a WT background. 0.01% corresponds to 1-2 copies of mutant DNA. For sensitivties of other KRAS Exon 2 hotspot mutations, please refer to Riva et al. 2017.
PlentiPlex™ MYD88 L265P assay is intended for in vitro diagnosis of the leucine to proline mutation in codon 265 of the Myeloid differentiation primary response 88 protein (MYD88 L265P) in genomic DNA (gDNA) samples. The obtained results of the PlentiPlex™ MYD88 L265P assay are intended for assisting in the discrimination between patients with Lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (LPL/WM) and non-Hodgkin lymphoma.