The invention consists of a novel analytical method for the PCR-free molecular detection of SARS-Cov-2 (of the RNA genome) by PCR-free technology which integrates a single-step sample extraction and a detection by cooperative hybridization combined with electrochemiluminescence based transduction.
| Patent title | Innovative PCR-free biotechnologies based on ElectroChemiluminescence for the quantification of the whole genome for the early diagnosis of SARS-CoV-2 viral infections |
|---|---|
| Thematic area | Health |
| Ownership | PERSONAL GENOMICS, ALMA MATER STUDIORUM - UNIVERSITA' DI BOLOGNA |
| Inventors | Giovanni Valenti, Katarzyna Malgorzata Kwiatkowska, Sabrina Conoci, Paolo Garagnani, Francesco Paolucci, Luca Prodi |
| Filed on | 30 March 2022 |
Currently there are no methods that are able to carry out the molecular analysis of COVID (buffer), that is, to detect the presence of the pathogen's genome in the biological sample, without PCR amplification.
The on going technical problem concerns the molecular detection and quantification of the RNA genome nucleic acid of COVID-19 without resorting to the use of PCR. To date, there is a lack of a solid and rational technological path for the practical implementation of biotechnologies without PCR.
The present invention relates to a process for the detection of an analyte in an isolated sample. The analyte in question is a target nucleic acid. The process comprises five stages:
- Providing a sample containing at least one nucleic acid.
- Heat-treating the sample.
- Contacting the heat-treated sample with at least two single-stranded nucleic acid probes. Each of these single-stranded nucleic acid probes is complementary to a corresponding portion of the target nucleic acid.
- Adding an active luminophore.
- Determining the luminescence signal generated by the active luminophore.
Direct Applications: Rapid Molecular Test for COVID 19 to be used for massive population screening.
Indirect applications: Rapid molecular testing for other RNA viruses and RNA genetic markers (e.g. miRNA).
Benefits:
• Molecular test (swab) of COVID without amplification
• Integrated extraction-detection system
• rapid and direct molecular determination (<1h)
• High analytical performance (high signal to noise ratio; <0.1 cps / reaction)
• Versatile methodology applicable to different RNA pathogens, which can be detected during the same analytical process (for example determining the cause of a specific pneumonia possibility of implementing rapid screening tests thanks to the advantages of POCs (point-of-care)