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Related Experiment Videos

Nucleic acid sequence-based amplification methods to detect avian influenza virus.

Lok-Ting Lau1, Jill Banks, Rebecca Aherne

  • 1Department of Neuroscience, Peking University Health Science Center, Key Laboratory of Neuroscience (Peking University), Neuroscience Research Institute, Peking University, Ministry of Education, 38 Xue Yuan Road, Beijing 100083, China.

Biochemical and Biophysical Research Communications
|December 20, 2003
PubMed
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Highly pathogenic avian influenza virus (AIV) poses significant threats. Researchers developed rapid Nucleic Acid Sequence-Based Amplification (NASBA) diagnostic tests for avian influenza detection, ensuring swift identification and control.

Area of Science:

  • Veterinary Virology
  • Molecular Diagnostics
  • Public Health

Background:

  • Highly pathogenic avian influenza virus (AIV) causes severe economic losses and poses a zoonotic risk.
  • Subtypes H5 and H7 are particularly concerning due to their high pathogenicity.
  • Rapid diagnostic tools are crucial for controlling AIV outbreaks and preventing human transmission.

Purpose of the Study:

  • To develop and validate rapid Nucleic Acid Sequence-Based Amplification (NASBA) assays for avian influenza virus detection.
  • To detect specific AIV subtypes (H5, H7) and all known subtypes (H1-H15).
  • To provide a sensitive, specific, and rapid diagnostic method for AIV surveillance.

Main Methods:

  • Development of two NASBA assays: one targeting the haemagglutinin gene for H5/H7 subtypes, and another targeting the matrix gene for all subtypes.

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  • Optimization of nucleic acid isolation, amplification, and detection within a 6-hour timeframe.
  • Validation of assay performance through sensitivity, specificity, and cross-reactivity testing against relevant viruses.

    • Main Results:

    • Successful development of NASBA assays capable of detecting AIV subtypes H5 and H7, and all known subtypes (H1-H15).
    • Assays demonstrated high sensitivity (five to seven orders of magnitude dynamic range) and specificity, with no cross-reactivity.
    • The entire diagnostic procedure, from nucleic acid isolation to detection, was completed within 6 hours.
    • NASBA assay results showed strong correlation with traditional viral culture methods.

    Conclusions:

    • NASBA technology offers a rapid, sensitive, and specific method for detecting avian influenza viruses.
    • These assays can significantly aid in the early detection and surveillance of AIV, crucial for disease control and public health.
    • The developed diagnostic tools are valuable for veterinary and public health laboratories managing AIV outbreaks.