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Related Experiment Videos

High performance Nanogold-silver in situ hybridisation

G W Hacker1

  • 1Institute of Pathological Anatomy, Salzburg General State Hospital, University of Salzburg, Austria. g.hacher@lkasbg.gv.at

European Journal of Histochemistry : EJH
|September 5, 1998
PubMed
Summary

A new Streptavidin-Nanogold-Silver in situ hybridisation (ISH) method offers single gene copy sensitivity for detecting nucleic acid sequences. This technique provides a sensitive and accessible alternative for various diagnostic applications.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Conventional in situ hybridisation (ISH) requires high target nucleic acid copy numbers (10-50 per cell).
  • In situ PCR offers single-copy sensitivity but suffers from high false-negative/positive rates.
  • Existing methods often require specialized, expensive equipment, limiting accessibility.

Purpose of the Study:

  • To introduce a novel, accessible in situ hybridisation technique for detecting low copy number nucleic acid sequences.
  • To evaluate the sensitivity and applicability of Streptavidin-Nanogold-Silver ISH combined with tyramide signal amplification (TSA™/CARD).

Main Methods:

  • Utilized Streptavidin-Nanogold-Silver in situ hybridisation (ISH).
  • Employed tyramide signal amplification (TSA™), also known as catalysed reporter deposition (CARD).

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  • Applied the assay to paraffin sections and cytological preparations.

    • Main Results:

    • Achieved single gene copy sensitivity for DNA virus detection.
    • Demonstrated very high sensitivity for RNA detection.
    • The assay is easy to perform in routine laboratories without expensive equipment.

    Conclusions:

    • Streptavidin-Nanogold-Silver ISH with TSA™/CARD is a highly sensitive and accessible method for detecting low copy number nucleic acids.
    • This technique has broad applications in early viral infection detection, cancer gene identification, genetic disease diagnosis, and mRNA detection.