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oai:repisalud.isciii.es:20.500.12105/90962024-09-27T21:55:25Zcom_20.500.12105_2052com_20.500.12105_2051col_20.500.12105_19609

00925njm 22002777a 4500 dc Caramalho, Rita author Madl, Lisa author Rosam, Katharina author Rambach, Günter author Speth, Cornelia author Pallua, Johannes author Larentis, Thomas author Araújo, Ricardo author Alastruey-Izquierdo, Ana author Lass-Flörl, Cornelia author Lackner, Michaela author 2019-10-11 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. J Fungi (Basel). 2019 Oct 11;5(4). pii: E98. 10.3390/jof5040098 2309-608X 2309-608X Journal of fungi (Basel, Switzerland) 31614610 http://hdl.handle.net/20.500.12105/9096 HRM Activity marker Growth stage Large subunit ribosomal DNA Mitochondrial genes Mucormycosis Pan-Mucorales qPCR Seminested PCR Evaluation of a Novel Mitochondrial Pan-Mucorales Marker for the Detection, Identification, Quantification, and Growth Stage Determination of Mucormycetes