Ultra-sensitive detection of nucleic acid mutations for research and clinical use

What is superRCA?

superRCA is an ultra-sensitive and highly specific molecular amplification technology. It is used to detect very small amounts of DNA sequence variants, like cancer mutations, in patient tissue and blood samples.

superRCA offers effective detection of multiple targets simultaneously – so called multiplexing. The assay can be performed in most hematology laboratories with existing equipment, using well-established flow cytometry for read-out, enabling more accessible testing with shorter response times.

The convenient, non-intrusive liquid biopsy-format enables higher frequency of testing for improved patient monitoring and earlier detection of relapse.

In the clinic, superRCA can be used by the practicing clinician to monitor disease progress for adjusted treatment and personalized medicine. Our first application is in Acute Myeloid Leukemia (AML), offering single and multiplex testing for recurring tumor-specific mutations.

For researchers and pharmaceutical developers, our RUO kits offer a convenient and efficient toolbox for investigating numerous therapeutic areas to design companion diagnostics and advance personalized medicine.

superRCA offers a personal diagnostics platform for clinicians while providing an open learning system for researchers, partners and collaborations that allows new discoveries.

Ultra-sensitivity – detects 1 mutation in 100,000 wild types

superRCA can detect 1 mutation out of a 100 000 wild-type DNA molecules.

The unprecedented sensitivity of superRCA is achieved by two consecutive Rolling Circle Amplification (RCA) reactions. A first standard RCA step is followed directly by a subsequent “in situPadlock Probing and RCA step. As a result the target region is genotyped with high specificity, enumerated with higher precision.

superRCA method

An indepth introduction to superRCA

superRCA utilizes Rolling Circle Amplification (RCA) and Padlock probes in a novel way to achieve highly specific, ultra-sensitive detection of nucleic sequences. The method produces a relatively large, self-constrained structure – a superRCA structure – that can be directly analyzed by microscopy or automated using flow cytometry without the need for partitioning.

Flow cytometry with fluorescent labelling ensures that multiple targets can be analyzed simultaneously. As a result, the platform is extremely effective for patient-near, fast and cost-effective patient monitoring as well as companion diagnostics and pharmaceutical development .

DNA is extracted from the sample, either whole blood, bone marrow or tissue.

The DNA sequences of interest, known to be mutated in a patient’s malignant cells, are first enriched by a limited pre-PCR (~10 cycle) amplification.

The enriched sample then undergoes a ligase-mediated circularization of one strand.

The circularized strands containing the target region are then amplified by the first Rolling Circle Amplification  (RCA) step.

This is followed by Padlock Probe ligation and a highly-specific, second RCA step. During this step, the second RCA encircles the first RCA product to form large superRCA structures that can be analyze by flow cytometry.

The superRCA products can be scored as mutant- or wildtype-specific using fluorophore-labeled hybridization probes and recorded as individual, brightly fluorescent objects in a standard flow cytometer. For multiplex assays, analysis is similarly done by using multiple fluorophores and wavelengths.

Are you interested in more details about our technology platform superRCA? Please contact us.

Book a meetingEnquire about a collaboration

Why superRCA makes a difference

  • Unprecedented ultra-sensitivity of 1 mutation in 100 000 wild types due to the novel combination of the extremely high specificity of Padlock probes and RCA
  • Meets or exceeds currently available technologies like ddPCR, NGS and traditional liquid biopsy assays by 10-100 times
  • Outstanding performance on High GC% targets. Assessment of high GC content with the same sensitivity as low GC content – with minimum bias.
  • High multiplexity creates opportunities for effective research, monitoring therapies and drug resistance, sensitive multiplex detection of mutations for treatment selection (companion diagnostics), quantitative measurement for patient follow up and clinical phase drug development
  • Convenient and reliable analysis by well-established flow cytometry for speed by patient-near testing.
  • Other read-out and analysis methods including fluorescence microscopy
  • Speed and cost-effectiveness at high-throughput.
  • An integrated sample-to-answer system with robotic automation using standard lab equipment and well-established instrumentation, minimizes manual handling and is highly amenable to clinical practice
  • Currently available as research use only kit or as a customized service