An Isothermal Deoxyribozyme Sensor for Rapid Detection of Enteroviral RNA

Area of Science:

• Environmental Science
• Molecular Biology
• Biotechnology

Background:

• Enteric viruses cause significant waterborne infections due to their stability and low infectious dose.
• Current molecular diagnostics require thermal cycling and centralized labs, hindering use in resource-limited areas.
• Developing field-deployable diagnostics is crucial for effective environmental surveillance.

Purpose of the Study:

• To develop an isothermal biosensor for detecting enteric viral RNA.
• To enhance the sensor's sensitivity using isothermal RNA amplification.
• To validate the sensor's performance with environmental water samples.

Main Methods:

• Developed a split deoxyribozyme biosensor that reconstitutes upon binding conserved enteroviral RNA sequences.
• Utilized a fluorogenic substrate cleaved by the activated deoxyribozyme to generate a fluorescent signal.
• Coupled the deoxyribozyme assay with isothermal RNA amplification for increased sensitivity.

Main Results:

• The biosensor demonstrated sequence-specific detection of enteroviral RNA.
• Successful validation was achieved using both synthetic RNA and RNA from environmental water samples.
• The amplified DNAzyme assay showed high selectivity and compatibility with real-world samples.

Conclusions:

• The developed DNAzyme-based platform offers a portable and sequence-specific method for enteric virus detection.
• Isothermal amplification significantly enhances the sensitivity of the viral RNA detection assay.
• This technology holds promise for field-deployable environmental surveillance of waterborne enteric viruses.