Ultrasensitive protein-level detection for respiratory infectious viruses

Area of Science:

• Biotechnology
• Infectious Diseases
• Molecular Diagnostics

Background:

• Influenza virus, adenovirus, and respiratory syncytial virus cause significant respiratory illnesses with overlapping symptoms.
• Current PCR diagnostics have limitations including cost, false negatives, and inability to differentiate live/dead viruses.
• There is a need for alternative protein-focused diagnostic methods for respiratory infections.

Purpose of the Study:

• To develop and validate a novel, sensitive, and cost-effective diagnostic assay for common respiratory viruses.
• To evaluate the performance of TN-cyclon™ in detecting influenza A, influenza B, adenovirus, and RSV.
• To establish the limit of detection (LOD) and sensitivity of the new assay.

Main Methods:

• Development of TN-cyclon™, an enzyme-linked immunosorbent assay utilizing thio-nicotinamide adenine dinucleotide cycling for signal amplification.
• Quantification of viral load for influenza A, influenza B, adenovirus, and RSV using the developed assay.
• Determination of LOD in moles/assay and plaque-forming units/mL (pfu/mL) for each virus type.

Main Results:

• TN-cyclon™ detected extremely low viral levels with LODs as low as 2.36 × 10^-18 moles/assay for adenovirus.
• The assay successfully detected viruses diluted in extract buffer at clinically relevant concentrations (e.g., 3 pfu/mL for influenza A).
• Significant differences were observed between viral samples and blank controls across all tested viruses.

Conclusions:

• TN-cyclon™ is a highly sensitive, low-cost, and user-friendly diagnostic technique for respiratory viruses.
• This novel assay offers a promising alternative to current PCR methods for diagnosing respiratory infections.
• The method demonstrates potential for rapid and accurate identification of influenza, adenovirus, and RSV.