Detection of Fusarium Species in Clinical Specimens by Probe-Based Real-Time PCR

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.

Trans-kingdom fungal pathogens infecting both plants and humans, and the problem of azole fungicide resistance

“…A broad range of Fusarium species can be detected with a probe-based real-time PCR assay. Springer et al ( 2019 ) designed a 28S-based analysis utilizing a specific hydrolysis probe which can be applied for the diagnosis of fusarioses from bronchoalveolar lavage fluid samples. Another PCR-based method employing a special instrument with electrochemical detection system was developed by Zhang et al ( 2020 ) for the identification of 15 fungal pathogens directly from blood cultures.…”

Mold detection and environmentally friendly prevention technology for animal specimens

“…fi xed, paraffi n-embedded tissue samples, Comparison of specifi c and broad-range fungal qPCR assays [16].…”

CRISPR-Cas12a-Based Diagnostics of Wheat Fungal Diseases

“…Morphological scanning can be used in-field and proceeded rapidly, but it is difficult to distinguish symptoms caused by different pathogens . Alternatively, DNA sequence-based molecular techniques have been extensively adopted in fungal species identification, such as polymerase chain reaction (PCR) using species-specific primers and relative quantification real-time PCR (qPCR). − Recently, droplet digital PCR (ddPCR) and loop-mediated isothermal amplification (LAMP) have become powerful tools for increasing the specificity and sensitivity of detection. ,− However, the nucleic acid amplification may result in nonspecific amplification, , adversely affecting the diagnostic accuracy of wheat FHB.…”