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Detection of SARS-CoV-2 Virus by Triplex Enhanced Nucleic Acid Detection Assay (TENADA)
Anna Aviñó1,2, Carlos Cuestas-Ayllón2,3, Manuel Gutiérrez-Capitán4
1Institute for Advanced Chemistry of Catalonia (IQAC), Consejo Superior de Investigaciones Científicas (CSIC), 08034 Barcelona, Spain.
International Journal of Molecular Sciences
|December 11, 2022
Summary
A new assay, triplex enhanced nucleic acid detection assay (TENADA), offers a rapid and sensitive method for detecting pathogens like SARS-CoV-2. This oligonucleotide hybridization technique provides accurate COVID diagnosis without amplification, enabling faster results.
Area of Science:
- Molecular Biology
- Biotechnology
- Nanotechnology
Background:
- Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) causes COVID-19, necessitating rapid diagnostic methods.
- Current Polymerase Chain Reaction (PCR) diagnostics are time-consuming and require specialized equipment.
- Immunological assays offer speed but may lack specificity.
Purpose of the Study:
- To develop an innovative, rapid, and sensitive diagnostic assay for pathogen detection.
- To adapt oligonucleotide hybridization for various biosensing platforms.
- To establish a nucleic acid detection method that bypasses amplification steps.
Main Methods:
- Utilized polypurine reverse-Hoogsteen hairpins (PPRHs) for high-affinity triplex formation with target nucleic acids.
- Employed a sandwich oligonucleotide hybridization approach for pathogen genome capture.
- Integrated labeled oligonucleotides for trimolecular complex detection, analogous to antigen tests.
Main Results:
- Achieved a limit of detection around 0.01 nM (femtomole levels) without amplification.
- Demonstrated adaptability to diverse biosensing devices: thermal lateral flow, electrochemical, and fluorescent microarrays.
- Developed the triplex enhanced nucleic acid detection assay (TENADA).
Conclusions:
- TENADA provides a sensitive and rapid detection method for viral genomes, including SARS-CoV-2.
- The assay's design allows for broad applicability to any pathogen with a known genome sequence.
- This approach offers a promising alternative to existing diagnostic methods, adaptable to various biosensing platforms.
