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Published on: February 23, 2018
Detection of Fusarium Species in Clinical Specimens by Probe-Based Real-Time PCR
Jan Springer1, Grit Walther2, Volker Rickerts3
1Department for Internal Medicine II, University Hospital of Wuerzburg, 97080 Wuerzburg, Germany.
Abstract:
The mold Fusarium is a ubiquitous fungus causing plant, animal and human infections. In humans, Fusarium spp. are the major cause of eye infections in patients wearing contact lenses or after local trauma. Systemic infections by Fusarium spp. mainly occur in immunosuppressed patients and can disseminate throughout the human body. Due to high levels of resistance to antifungals a fast identification of the causative agent is an urgent need. By using a probe-based real-time PCR assay specific for the genus Fusarium we analysed several different clinical specimens detecting Fusarium spp. commonly found in clinical samples in Germany. Also, a large collection of lung fluid samples of haematological patients was analysed (n = 243). In these, two samples (0.8%) were reproducibly positive, but only one could be confirmed by sequencing. For this case of probable invasive fungal disease (IFD) culture was positive for Fusarium species. Here we describe a rapid, probe-based real-time PCR assay to specifically detect DNA from a broad range of Fusarium species and its application to clinically relevant specimens.
Insights
A new real-time PCR assay rapidly detects Fusarium species, a fungus causing infections in humans. This diagnostic tool is crucial for identifying the causative agent, especially with increasing antifungal resistance.
Area of Science:
- Medical Mycology
- Molecular Diagnostics
- Infectious Diseases
Background:
- Fusarium species are ubiquitous fungi causing infections in plants, animals, and humans.
- Human infections include eye infections and systemic infections, particularly in immunosuppressed individuals.
- High antifungal resistance necessitates rapid identification of Fusarium species.
Purpose of the Study:
- To develop and evaluate a rapid, probe-based real-time PCR assay for specific Fusarium genus detection.
- To assess the assay's performance on various clinical specimens, including lung fluid from hematological patients.
Main Methods:
- Development of a probe-based real-time PCR assay targeting the Fusarium genus.
- Analysis of clinical specimens, including 243 lung fluid samples from hematological patients.
- Confirmation of positive results using DNA sequencing and fungal culture.
Main Results:
- The real-time PCR assay successfully detected Fusarium species in clinical samples from Germany.
- Two out of 243 (0.8%) lung fluid samples were positive, with one confirmed by sequencing and culture.
- Culture confirmed one case of probable invasive fungal disease (IFD) caused by Fusarium species.
Conclusions:
- A rapid, probe-based real-time PCR assay can effectively detect a broad range of Fusarium species.
- This assay is a valuable tool for the rapid identification of Fusarium in clinical settings.
- Early and accurate diagnosis is critical for managing Fusarium infections, especially in light of antifungal resistance.
