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BRET-based biosensors for SARS-CoV-2 oligonucleotide detection
Asfia Sultana1, Anupriya M Geethakumari1, Zeyaul Islam2
1Division of Biological and Biomedical Sciences, College of Health and Life Sciences, Hamad Bin Khalifa University, Education City, Qatar Foundation, Doha, Qatar.
Frontiers in Bioengineering and Biotechnology
|June 18, 2024
Summary
Researchers developed a novel bioluminescence resonance energy transfer (BRET)-based biosensor for oligonucleotide detection (BioOD). This rapid, sensitive, and cost-effective tool enables specific detection of SARS-CoV-2 variants at the point-of-care.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Virology
Background:
- The emergence of novel viruses and variants necessitates rapid point-of-care diagnostic tools.
- Existing nucleic acid amplification methods like RT-qPCR and LAMP have limitations in ease of use and robustness.
- A facile and sensitive platform is required for early detection of viral pathogens.
Purpose of the Study:
- To engineer a versatile, sensitive, rapid, and cost-effective bioluminescence resonance energy transfer (BRET)-based biosensor for oligonucleotide detection (BioOD).
- To demonstrate the potential of BioODs for specific detection of SARS-CoV-2 variants.
- To validate the BioOD platform as a proof-of-principle for point-of-care diagnostics.
Main Methods:
- Designed BioODs targeting SARS-CoV-2 parental and Delta variants using fluorescently modified molecular beacons and NanoLuc luciferase.
- Conjugated sensor modules via a complementary oligonucleotide handle DNA.
- Assessed BioODs for kinetics, affinity, specificity, and signal detection via color change in cell phone images.
Main Results:
- The developed BioODs demonstrated rapid and highly specific detection of viral oligonucleotides.
- A decrease in BRET efficiency, indicated by a spectral shift, correlated with oligonucleotide binding.
- Signal changes were visually detectable as color shifts using cell phone camera images.
Conclusions:
- The engineered BioODs offer a promising platform for sensitive and specific oligonucleotide detection.
- This technology has potential applications in point-of-care testing for viral infections, including variant specificity.
- BioODs could significantly aid in large-scale viral infection surveillance and early outbreak detection.
