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In Vitro Selection of Aptamers to Differentiate Infectious from Non-Infectious Viruses
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Highly-Efficient Selection of Aptamers for Quantitative Fluorescence Detecting Multiple IAV Subtypes.

Meng Wang1, Jianjun Chen2,3, Zhi-Ling Zhang1

  • 1College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, Hubei 430072, China.

Analytical Chemistry
|September 11, 2024
PubMed
Summary

Rapid detection of multiple influenza A virus (IAV) subtypes is now possible. This new method uses aptamers for sensitive, quantitative fluorescence detection of H5N1, H7N9, and H9N2 strains within an hour.

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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.