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Updated: Jul 15, 2026
High-throughput Detection Method for Influenza Virus
Published on: February 4, 2012
Ultrasensitive protein-level detection for respiratory infectious viruses
Yuki Kobayashi1,2, Yuta Kyosei1, Ryutaro Ogawa1
1Department of Biology, Waseda University, Tokyo, Japan.
Abstract:
Influenza virus, adenovirus, and respiratory syncytial virus cause major respiratory infections. These infections have similar initial symptoms making it difficult to differentiate them based on symptoms alone. PCR is currently used as the standard diagnostic test for these infections, however, it has its limitations such as non-specific and false-negative amplifications, high cost, and the inability to distinguish between a live or dead virus. Therefore, there is a need for alternative diagnostic methods that focus on protein. Here, we introduce TN-cyclon™, which is an enzyme-linked immunosorbent assay combined with thio-nicotinamide adenine dinucleotide cycling to amplify signals, rather than the protein itself. Using this method, we were able to detect extremely low levels of viruses such as influenza A, influenza B, adenovirus, and RS virus, with LODs of 2.96 × 10-18 moles/assay, 2.98 × 10-18 moles/assay, 2.36 × 10-18 moles/assay, and 3.55 × 10-18 moles/assay, respectively. Furthermore, we successfully detected viruses diluted with extract buffer, with a significant difference to the blank at concentrations of 3 pfu/mL for influenza A, 1000 pfu/mL for influenza B, 43.8 pfu/mL for adenovirus, and 125 pfu/mL for RS virus. This shows that our low-cost and easy-to-use technique has sufficient sensitivity in diagnosing respiratory infections.
Insights
A new diagnostic method, TN-cyclon™, offers a sensitive and low-cost way to detect respiratory viruses like influenza and RSV. This enzyme-linked immunosorbent assay overcomes limitations of PCR for faster, more accurate respiratory infection diagnosis.
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.
