About our ASPYRE technology

ASPYRE is a completely new, highly sensitive method of identifying actionable genetic information in biological samples to inform the diagnosis, treatment, and monitoring of disease.

At the heart of the technology is an enzymatic reaction called pyrophosphorolysis, in which a DNA polymerase is forced to run in reverse, digesting double-stranded DNA rather than synthesising it.

Crucially, this reaction only proceeds through perfectly matched double-stranded DNA and is blocked by mismatches.

 

How does ASPYRE work?

  • First, DNA is isolated from tissue samples or blood plasma, and target genes are enriched through amplification. In our ASPYRE-Lung assay this enrichment covers all 11 genes recommended for testing in NCCN guidelines for precision lung cancer therapy.
  • We then add a panel of synthetic barcoded DNA probes, designed to target more than 100 specific actionable mutations across these target genes.
  • The probes hybridize with the target regions from the patient sample. They are perfectly matched to any mutated sequences that are present, but will be mismatched against healthy DNA and non-target sequences.
Probe hybridization
 
 

Next, the pyrophosphorolysis reaction selectively digests only the perfectly matched probes, releasing the probe tail. Probes that have hybridised to healthy DNA are only digested to the point of mismatch, leaving an extended tail.

Probe digestion and tail release
 
 

The probe tails are then circularized by joining the two ends together. Only fully digested probes released from target sequences can be joined into a perfect circle, while probes released from non-target sequences or healthy DNA cannot.

Probe tail circularization
 
 

Finally, perfectly circularized probes are amplified and detected via their unique barcodes, revealing which target mutations were present in the patient sample.

Probe tail amplification and detection
 
 

Key benefits of ASPYRE

  • Ultra-sensitive: extreme specificity of the pyrophosphorolysis reaction enables single molecule sensitivity, providing answers from small samples with low tumor content
  • Scalable: amplification of synthetic probes, not sample DNA, enables detection of hundreds of markers across multiple genes in a single assay, without compromising performance
  • Faster, cheaper and more targeted than NGS: <4 hours to actionable information at a fraction of the cost
  • More informative than PCR-based approaches: avoid wasting precious sample by analyzing more than 100 markers in a single assay, with best-in-class sensitivity
  • Simple workflow requiring no new instrumentation: runs on existing PCR equipment
 
 

Find out more

Learn more about ASPYRE from our scientific publications: