One of the main challenges impeding genomic-based disease identification and characterization stems from the fact that related analytes, such as pathogens genes and cancer biomarkers, are present in extremely low concentrations in biomedical samples. Thus, currently, most clinical genomic identification approaches are based on amplification methods, such as PCR (Polymerase Chain Reaction). Yet this introduces high error values to any diagnosis, involves expensive reagents, and generally restricts the level of multiplexity of the test and hence its accuracy. While next-generation sequencing methods allow for whole-genome sequencing, this method requires lengthy sample preparation, often involving amplification as well, and yields much more data than is necessary for accurate classification of DNA and thus requires extensive processing and analysis. While the ligation reaction is a common method used for SNV detection due to its high specificity, low cost, and simplicity, such techniques suffer from low sensitivity due to the low signal to noise ratio of detection methods and limited ability for multiplexing.
The present patented technology is a ssNP-based, genotyping technique that circumvents DNA amplification and sequencing, and can provide unprecedented accuracy in identification of genetic variations such as SNVs and insertions/deletions in extremely short time. The invention involves the creation of an assay to uniquely transform genetic variations to molecular forms that are readily observed by nanopores at the single molecule level. The signal acquired by the nanopore is interpreted as molecularly encoded barcodes, which allow for unique classification of the source genome with high diagnostic accuracy.
- Combines the specificity of a ligation reaction with the ultra-sensitivity of nanopore biosensors
- Quick & Accurate
- Allows for multiplexed detection
Applications and Opportunities
- Pathogen diagnosis
- Early-stage cancer detection based on circulating tumor DNA