Highly-Efficient Selection of Aptamers for Quantitative Fluorescence Detecting Multiple IAV Subtypes

Area of Science:

• Virology
• Biotechnology
• Biosensing

Background:

• Influenza A virus (IAV) poses significant threats to human and animal health.
• Rapid mutation of IAVs necessitates advanced detection methods for various subtypes.
• Simultaneous detection of multiple IAVs is critical for effective disease control.

Purpose of the Study:

• To develop a sensitive and quantitative method for simultaneous detection of multiple IAV subtypes.
• To screen and validate specific aptamers targeting hemagglutination (HA) proteins of H5N1, H7N9, and H9N2 viruses.
• To enable rapid, fluorescence-based quantitative detection of influenza A viruses.

Main Methods:

• Screening of specific aptamers against HA proteins of H5N1, H7N9, and H9N2 using a multichannel magnetic microfluidic chip.
• Truncation and labeling of aptamers with distinct fluorescence markers.
• Development of an aptamer sandwich assay for quantitative fluorescence detection.

Main Results:

• Aptamers with nanomolar affinity and high specificity for H5N1, H7N9, and H9N2 HA proteins were identified.
• The developed aptamer sandwich assay achieved quantitative fluorescence detection within 1 hour.
• Low limits of detection were achieved: 0.38 TCID50/mL (H5N1), 0.75 TCID50/mL (H7N9), and 1.14 TCID50/mL (H9N2).
• The method demonstrated excellent specificity, anti-interference capabilities, and reproducibility.

Conclusions:

• A sensitive and quantitative fluorescence detection method for H5N1, H7N9, and H9N2 influenza A viruses was successfully developed.
• This method enables the simultaneous, rapid, and quantitative detection of multiple IAV subtypes.
• The findings contribute to improved diagnostics and surveillance of influenza A virus infections.