Next Article in Journal / Special Issue
First Isolation, Antifungal Susceptibility, and Molecular Characterization of Cryptococcus neoformans from the Environment in Croatia
Previous Article in Journal
Antifungal Drugs: Special Problems Treating Central Nervous System Infections
Previous Article in Special Issue
Identification of Mycoses in Developing Countries
Open AccessArticle

Evaluation of a Novel Mitochondrial Pan-Mucorales Marker for the Detection, Identification, Quantification, and Growth Stage Determination of Mucormycetes

1
Institute of Hygiene and Medical Microbiology, Medical University of Innsbruck, Schöpfstraße 41, 6020 Innsbruck, Austria
2
Institute of Pathology, Neuropathology and Molecular Pathology, Medical University of Innsbruck, Müllerstraße 44, 6020 Innsbruck, Austria
3
i3 S, Instituto de Investigacao e Inovacao da Universidade do Porto, R. Alfredo Allen, 4200-135 Porto, Portugal
4
Department of Medical Biotechnology, School of Health Sciences, Flinders University, Sturt Rd, Bedford Park SA 5042, Australia
5
Mycology Reference Laboratory, National Centre for Microbiology. Instituto de Salud Carlos III. Majadahonda, 28220 Madrid, Spain
*
Author to whom correspondence should be addressed.
These authors contributed equally to the paper.
J. Fungi 2019, 5(4), 98; https://doi.org/10.3390/jof5040098
Received: 20 August 2019 / Revised: 30 September 2019 / Accepted: 8 October 2019 / Published: 11 October 2019
(This article belongs to the Special Issue Molecular Diagnostics of Fungal Infections)
Mucormycosis infections are infrequent yet aggressive and serious fungal infections. Early diagnosis of mucormycosis and its discrimination from other fungal infections is required for targeted treatment and more favorable patient outcomes. The majority of the molecular assays use 18 S rDNA. In the current study, we aimed to explore the potential of the mitochondrial rnl (encoding for large-subunit-ribosomal-RNA) gene as a novel molecular marker suitable for research and diagnostics. Rnl was evaluated as a marker for: (1) the Mucorales family, (2) species identification (Rhizopus arrhizus, R. microsporus, Mucor circinelloides, and Lichtheimia species complexes), (3) growth stage, and (4) quantification. Sensitivity, specificity, discriminatory power, the limit of detection (LoD), and cross-reactivity were evaluated. Assays were tested using pure cultures, spiked clinical samples, murine organs, and human paraffin-embedded-tissue (FFPE) samples. Mitochondrial markers were found to be superior to nuclear markers for degraded samples. Rnl outperformed the UMD universal® (Molyzm) marker in FFPE (71.5% positive samples versus 50%). Spiked blood samples highlighted the potential of rnl as a pan-Mucorales screening test. Fungal burden was reproducibly quantified in murine organs using standard curves. Identification of pure cultures gave a perfect (100%) correlation with the detected internal transcribed spacer (ITS) sequence. In conclusion, mitochondrial genes, such as rnl, provide an alternative to the nuclear 18 S rDNA genes and deserve further evaluation. View Full-Text
Keywords: mucormycosis; HRM; qPCR; large subunit ribosomal DNA; mitochondrial genes; growth stage; activity marker; seminested PCR; pan-Mucorales mucormycosis; HRM; qPCR; large subunit ribosomal DNA; mitochondrial genes; growth stage; activity marker; seminested PCR; pan-Mucorales
Show Figures

Figure 1

MDPI and ACS Style

Caramalho, R.; Madl, L.; Rosam, K.; Rambach, G.; Speth, C.; Pallua, J.; Larentis, T.; Araujo, R.; Alastruey-Izquierdo, A.; Lass-Flörl, C.; Lackner, M. Evaluation of a Novel Mitochondrial Pan-Mucorales Marker for the Detection, Identification, Quantification, and Growth Stage Determination of Mucormycetes. J. Fungi 2019, 5, 98.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop