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Differential Diagnosis of Fungal Pneumonias vs. Tuberculosis in AIDS Patients by Using Two New Molecular Methods

Leticia Bernal-Martínez1, Laura Herrera2, Clara Valero1

  • 1Centro Nacional de Microbiología, Mycology Reference Laboratory, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.

Journal of Fungi (Basel, Switzerland)
|April 30, 2021
PubMed

Insights

New PCR assays offer rapid differential diagnosis for opportunistic fungal pneumonias and tuberculosis in AIDS patients. These sensitive and reproducible methods improve early detection, crucial for timely treatment and better patient outcomes.

Area of Science:

  • Medical Diagnostics
  • Infectious Diseases
  • Molecular Biology

Background:

  • Opportunistic fungal pneumonias (OFP) are a leading cause of mortality in AIDS patients globally.
  • Diagnosis is often delayed due to initial suspicion of tuberculosis (TB), compounded by limitations and inaccessibility of current diagnostic tools in resource-limited settings.

Purpose of the Study:

  • To develop and validate novel molecular assays for the differential diagnosis of common fungal pathogens (Histoplasma capsulatum, Cryptococcus neoformans/C. gattii, Pneumocystis jirovecii) causing OFP versus Mycobacterium tuberculosis complex (MTBC) in AIDS patients.

Main Methods:

  • Development and in vitro optimization of two real-time PCR-based assays: a multiplex real-time PCR (MRT-PCR) and a melting curve analysis PCR (MC-PCR).
  • Assays were validated using 42 clinical samples from patients with proven fungal infections or TB, alongside 33 controls.

Main Results:

  • Both MRT-PCR and MC-PCR demonstrated high reproducibility (CVs 1.84–4.83%), 100% specificity, and excellent detection limits (2–20 fg DNA).
  • The overall sensitivity was 88% for MRT-PCR and 90.4% for MC-PCR.
  • Both techniques proved to be fast, sensitive, and reproducible for pathogen detection and differential diagnosis.

Conclusions:

  • The developed MRT-PCR and MC-PCR assays are effective tools for the rapid and accurate differential diagnosis of OFP and TB in AIDS patients.
  • These assays address the limitations of current diagnostic methods, offering potential for improved patient management, especially in resource-limited regions.
Keywords:
differential diagnosisfungal pneumoniatuberculosis

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