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Related Experiment Videos

In situ hybridization in paracoccidioidomycosis.

T De Brito1, G S Sandhu, B C Kline

  • 1Institute of Tropical Medicine and School of Dentistry, S. Paulo University, SP, Brazil. thalesbr@usp.br

Medical Mycology
|July 28, 1999
PubMed
Summary

In situ hybridization (ISH) successfully detected Paracoccidioides brasiliensis fungal cells in patient biopsies and guinea pig tissues. The study optimized probe sequences for improved fungal cell identification in paracoccidioidomycosis research.

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Area of Science:

  • Mycology
  • Infectious Diseases
  • Molecular Biology

Background:

  • Paracoccidioidomycosis is a systemic fungal infection caused by Paracoccidioides brasiliensis.
  • Accurate detection of the fungus in host tissues is crucial for diagnosis and understanding pathogenesis.
  • In situ hybridization (ISH) offers a method for visualizing specific microbial targets within tissue samples.

Purpose of the Study:

  • To evaluate the efficacy of in situ hybridization (ISH) using specific oligoprobes for detecting Paracoccidioides brasiliensis.
  • To compare the labeling efficiency of a specific probe and its complementary sequence in infected tissues.

Main Methods:

  • Oral biopsies from paracoccidioidomycosis patients and inoculated guinea pig testes were analyzed.
  • In situ hybridization (ISH) was performed using a 14 base-pair specific oligoprobe and its complementary sequence.

Related Experiment Videos

  • Combined ISH with Gridley stain for fungal cell wall visualization.

    • Main Results:

    • Approximately 2-3% of fungal cells were labeled when using the specific oligoprobe in conjunction with the Gridley stain.
    • Utilizing the complementary probe resulted in higher labeling efficiency, reaching up to 3% of fungal cells.
    • ISH demonstrated the presence of P. brasiliensis in both human and animal tissue models.

    Conclusions:

    • In situ hybridization (ISH) is a viable method for detecting Paracoccidioides brasiliensis in clinical and experimental settings.
    • Optimization of probe sequences, such as using complementary probes, can enhance the sensitivity of fungal detection.
    • This technique aids in the study of paracoccidioidomycosis and the distribution of the fungus within host tissues.