RNA polymerization actuating nucleic acid membrane (RANAM)-based biosensing for universal RNA virus detection

Area of Science:

• Biotechnology
• Nanotechnology
• Molecular Diagnostics

Background:

• Early diagnosis of viral infections like COVID-19 is critical for controlling spread and reducing mortality.
• Traditional diagnostic methods can be time-consuming, hindering rapid response during pandemics.
• Developing swift and precise diagnostic tools for RNA viruses remains a significant challenge.

Purpose of the Study:

• To develop a novel, rapid, and non-PCR-based colorimetric biosensor for RNA virus detection.
• To utilize an RNA polymerization actuating nucleic acid membrane (RANAM) for sensing RNA polymerase activity.
• To demonstrate the potential of this technology for early viral infection diagnosis.

Main Methods:

• A gold-metallized RNA polymerization actuating nucleic acid membrane (RANAM) was employed for colorimetric detection.
• The biosensor detects RNA polymerase activity by observing the inhibition of a redox reaction involving TMB and Au3+.
• Viral RNA-dependent RNA polymerase (RdRP) was used as the target molecule for proof-of-concept.

Main Results:

• The RANAM biosensor detected RNA-dependent RNA polymerase (RdRP) activity within 10 minutes.
• The prototype kit demonstrated high sensitivity, detecting RdRP at concentrations as low as 100 aM.
• Smartphone-assisted color imaging enabled simple and clear confirmation of results.

Conclusions:

• This novel RANAM-based biosensor offers a rapid, sensitive, and non-PCR method for detecting RNA viruses and their variants.
• The technology has potential for developing easy-to-use diagnostic kits for timely viral infection management.
• The colorimetric sensing strategy provides a visual readout for quick diagnosis.