Diagnostic approaches for dimorphic fungi

Diagnostic approaches for dimorphic fungi

Diagnostic approaches for dimorphic fungi

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Specimen & Microscopy - First Look

  • Specimens: Sputum, bronchoalveolar lavage (BAL), tissue biopsy, CSF, blood.
  • Stains: KOH prep (rapid), Giemsa, Gomori methenamine silver (GMS), and Periodic acid-Schiff (PAS). GMS & PAS are superior for tissue visualization.
  • Direct Visualization: Identify the characteristic yeast form in tissue samples (at 37°C).
    • Blastomyces: Broad-based budding yeast.
    • Coccidioides: Spherules containing endospores.
    • Histoplasma: Small, oval yeast, often within macrophages.
    • Sporothrix: Cigar-shaped or pleomorphic yeast.

GMS stain of dimorphic fungi yeast forms

Histoplasma capsulatum yeast forms are very small (2-5 µm) and characteristically found intracellularly within macrophages, distinguishing them from the larger Blastomyces yeast.

📌 Yeast in the heat, mold in the cold.

Fungal Culture - The Gold Standard

  • Definitive diagnosis by demonstrating thermal dimorphism.
  • Procedure:
    • Grown on Sabouraud Dextrose Agar (SDA).
    • Incubated at two temperatures:
      • 25-30°C: Grows as filamentous mold (saprophytic/environmental form).
      • 37°C: Converts to yeast form (parasitic/tissue form).
  • Limitation:
    • Slow growth, often taking 2-4 weeks for results.
  • ⚠️ Warning: Mold forms are highly infectious (airborne spores); require Biosafety Level (BSL)-3 precautions.

⭐ The ability to switch between mold and yeast forms based on temperature is the hallmark of dimorphic fungi. Remember the mnemonic: "Mold in the Cold, Yeast in the Heat/Beast."

Dimorphic fungus culture: mold at 25C, yeast at 37C

Serology & Antigen Tests - Rapid Results

  • Antibody (Ab) Detection

    • Methods: Immunodiffusion (ID), Complement Fixation (CF).
    • Detects host immune response (IgM, IgG).
    • Utility: Primarily for chronic pulmonary or disseminated disease (e.g., Coccidioidomycosis).
    • Limitations: Slow (weeks for seroconversion), cross-reactivity, low sensitivity in immunocompromised.
  • Antigen (Ag) Detection

    • Methods: Enzyme Immunoassay (EIA) on urine, serum, or CSF.
    • Utility: Rapid diagnosis, especially in acute/disseminated cases.
    • High-yield tests:
      • Histoplasma urinary Ag: High sensitivity in disseminated/progressive disease.
      • Blastomyces urinary Ag: Good sensitivity, but cross-reacts with Histoplasma.

High-Yield: The Histoplasma capsulatum urinary antigen test is highly sensitive (>90%) for disseminated histoplasmosis, particularly in immunocompromised patients, and can be used to monitor response to therapy as antigen levels decline with successful treatment.

ELISA Assay Principle for Antigen Detection

Histopathology & PCR - Tissue & Tech

  • Histopathology: Biopsy provides definitive diagnosis.
    • Stains: Gomori methenamine-silver (GMS) and periodic acid-Schiff (PAS) highlight fungal elements.
    • Identify characteristic tissue-phase shapes (yeast, spherules).
  • PCR: Amplifies fungal DNA for rapid identification.
    • High sensitivity and specificity.
    • Useful for culture-negative samples or sterile sites.

Paracoccidioides brasiliensis yeast forms show classic "captain's wheel" or "mariner's wheel" budding on microscopy, a high-yield visual for exams.

Paracoccidioides brasiliensis: captain's wheel budding

High‑Yield Points - ⚡ Biggest Takeaways

  • Dimorphism is the cornerstone: remember “yeast in the beast, mold in the cold.”
  • Culture is the gold standard for diagnosis but is slow and biohazardous.
  • Direct microscopy with special stains (GMS, PAS) on tissue is a rapid, key step.
  • Urine antigen testing is highly sensitive and specific for disseminated Histoplasmosis.
  • Blastomycosis is identified by its characteristic broad-based budding yeast.
  • Coccidioidomycosis is diagnosed by finding spherules containing endospores in tissue.

Practice Questions: Diagnostic approaches for dimorphic fungi

Test your understanding with these related questions

The surgical equipment used during a craniectomy is sterilized using pressurized steam at 121°C for 15 minutes. Reuse of these instruments can cause transmission of which of the following pathogens?

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Flashcards: Diagnostic approaches for dimorphic fungi

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Blastomycosis are _____ RBCs

TAP TO REVEAL ANSWER

Blastomycosis are _____ RBCs

the same size as (larger than/smaller than/the same size as)

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