PentaBase ApS – SensiScreen®
PentaBase ApS is a privately held company founded in 2006, having roots back to the invention Intercalating Nucleic Acid, INA®, made by the founder of PentaBase, Ulf B. Christensen and published in 2002 [Christensen UB and Pedersen EB, Nucl. Acid. Res., 2002, 30, 4918-4925]. Today PentaBase has refined INA®, and made it compatible with standard, open platform instruments used in most molecular biology facilities around the world. We call the proprietary monomers in our updated INA for “pentabases”.
（PentaBase’s website is here）
―Assays for Sensitive Detection of Cancer Mutations―
SensiScreen® are real-time PCR-based assays for somatic mutation detection in cancer patients. SensiScreen® combines high sensitivity with multiplexing capabilities and ease-of-use, and are designed to work on standard real-time PCR equipment. Sensitive detection of somatic mutations by SensiScreen® is achieved with PentaBase’s novel and selective INA® technologies including the use of BaseBlockers™ that suppress false positive signals from wild type templates.
●High sensitivity – down to one copy detection
●Simplex and multiplex configurations
●Detection in solid and liquid biopsies
●Open platform designs
From sample to answer in less than three hours
SensiScreen® 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 technology is 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® 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: 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: SensiScreen® FFPE assays have a limit of detection between 0.1% and 1% mutant DNA in a wild type background. SensiScreen® Liquid detects down to one copy of mutant DNA.
・Easy and fast: Easy to setup (hands-on-time <2 min/sample) and takes less than three hours from sample to result including DNA extraction.
・State of the art: Based on PentaBase’s proprietary INA® technology. Finds more true mutation positive samples compared to other common methods for somatic mutation testing (Riva et al. 2017).
・Proven: SensiScreen® CE IVD 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.
・Simplex and multiplex: Supplied in multiplex configurations for mutation screening and simplex configurations for genotyping and monitoring.
・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.
・Analysis of liquid biopsies
・Early detection and monitoring of cancer DNA biomarkers
・Detects down to 1 copy of mutated DNA
・From biopsy to result in less than 3 hours
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.
SensiScreen Liquid product list
|1701-1702||KRAS EXON2+3+4 Multiplex (K1)||12/60 reactions|
|1706-1707||KRAS EXON2 Multiplex (K2)||12/60 reactions|
|1711-1712||KRAS EXON3 Multiplex (K3)||12/60 reactions|
|1716-1717||KRAS EXON4 Multiplex (K4)||12/60 reactions|
|1721-1722||KRAS EXON2 Simplex (K5+K6)||12/60 reactions|
|1726-1727||KRAS EXON2 Simplex (K5)||12/60 reactions|
|1731-1732||KRAS EXON2 Simplex (K6)||12/60 reactions|
|1736-1737||KRAS EXON3 Simplex (K7+K8)||12/60 reactions|
|1741-1742||KRAS EXON3 Simplex (K7)||12/60 reactions|
|1746-1747||KRAS EXON3 Simplex (K8)||12/60 reactions|
|1751-1752||KRAS EXON4 Simplex (K9+K10)||12/60 reactions|
|1756-1757||KRAS EXON4 Simplex (K9)||12/60 reactions|
|1761-1762||KRAS EXON4 Simplex (K10)||12/60 reactions|
|1771-1772||NRAS EXON2+3+4 Multiplex (N1)||12/60 reactions|
|1776-1777||NRAS EXON2 Multiplex (N2)||12/60 reactions|
|1781-1782||NRAS EXON3 Multiplex (N3)||12/60 reactions|
|1786-1787||NRAS EXON4 Multiplex (N4)||12/60 reactions|
|1791-1792||NRAS EXON2 Simplex (N5+N6)||12/60 reactions|
|1796-1797||NRAS EXON2 Simplex (N5)||12/60 reactions|
|1801-1802||NRAS EXON2 Simplex (N6)||12/60 reactions|
|1806-1807||NRAS EXON3 Simplex (N7+N8)||12/60 reactions|
|1811-1812||NRAS EXON3 Simplex (N7)||12/60 reactions|
|1816-1817||NRAS EXON3 Simplex (N8)||12/60 reactions|
|1821-1822||NRAS EXON4 Simplex (N9)||12/60 reactions|
|1831-1832||BRAF V600 Multiplex (B1)||12/60 reactions|
|1836-1837||BRAF V600 Simplex (B2)||12/60 reactions|
|1841-1842||BRAF V600E Simplex (B3)||12/60 reactions|
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.
・Analysis of primary tumors and metastases
・Multiplex assays for screening of cancer mutation hotspots
・Simplex assays for genotyping and confirmation
・FFPE assays are applicable to fresh, frozen or FFPE biopsies
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.
SensiScreen FFPE product list
|1900-1901||KRAS EXON2+3+4 Multiplex||20/50 reactions|
|1905-1906||KRAS EXON2 Multiplex||20/50 reactions|
|1910-1911||KRAS EXON3 Multiplex||20/50 reactions|
|1915-1916||KRAS EXON4 Multiplex||20/50 reactions|
|1920-1921||KRAS EXON2 Simplex A+B||20/50 reactions|
|1925-1926||KRAS EXON2 Simplex A||20/50 reactions|
|1930-1931||KRAS EXON2 Simplex B||20/50 reactions|
|1935-1936||KRAS EXON3 Simplex A+B||20/50 reactions|
|1940-1941||KRAS EXON3 Simplex A||20/50 reactions|
|1945-1946||KRAS EXON3 Simplex B||20/50 reactions|
|1950-1951||KRAS EXON4 Simplex A+B||20/50 reactions|
|1955-1956||KRAS EXON4 Simplex A||20/50 reactions|
|1960-1961||KRAS EXON4 Simplex B||20/50 reactions|
|2000-2001||NRAS EXON2+3+4 Multiplex||20/50 reactions|
|2005-2006||NRAS EXON2 Multiplex||20/50 reactions|
|2010-2011||NRAS EXON3 Multiplex||20/50 reactions|
|2015-2016||NRAS EXON4 Multiplex||20/50 reactions|
|2020-2021||NRAS EXON2 Simplex A+B||20/50 reactions|
|2025-2026||NRAS EXON2 Simplex A||20/50 reactions|
|2030-2031||NRAS EXON2 Simplex B||20/50 reactions|
|2035-2036||NRAS EXON3 Simplex A+B||20/50 reactions|
|2040-2041||NRAS EXON3 Simplex A||20/50 reactions|
|2045-2046||NRAS EXON3 Simplex B||20/50 reactions|
|2050-2051||NRAS EXON4 Simplex||20/50 reactions|
|1398-1399||BRAF V600 Multiplex||20/50 reactions|
|1402-1403||BRAF V600 Simplex||20/50 reactions|
|1400-1401||BRAF V600E Simplex||20/50 reactions|
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