Spectrum of Zygomycete Species Identified in Clinically Significant Specimens in the United States

Abstract

Several members of the order Mucorales (subphylum Mucoromycotina) are important agents of severe human infections. The identification of these fungi by using standard mycologic methods is often difficult and time consuming. Frequently, the etiological agent in clinical cases is reported either as a Mucor sp., which is not the most frequent genus of zygomycetes, or only as a member of the Mucorales. For this reason, the actual spectrum of species of zygomycetes and their incidences in the clinical setting is not well known. The goals of this study were to compare the results of the molecular identification of an important set of clinical isolates, received in a mycological reference center from different regions of the United States, with those obtained by using the traditional morphological methods and to determine the spectrum of species involved. We tested 190 isolates morphologically identified as zygomycetes by using sequencing of the internal transcribed spacer (ITS) region of the ribosomal DNA. Molecular identification revealed that Rhizopus oryzae represented approximately half (44.7%) of these isolates. The remainder was identified as Rhizopus microsporus (22.1%), Mucor circinelloides (9.5%), Mycocladus corymbifer (formerly Absidia corymbifera) (5.3%), Rhizomucor pusillus (3.7%), Cunninghamella bertholletiae (3.2%), Mucor indicus (2.6%), Cunninghamella echinulata (1%), and Apophysomyces elegans (0.5%). The most common anatomic sites for clinically significant zygomycetes, as determined by isolates sent to the Fungus Testing Laboratory for identification and/or susceptibility testing and included in this study, were the sinuses, lungs, and various cutaneous locations, at 25.8%, 26.8%, and 28%, respectively. These sites represented approximately 80% of the isolates evaluated. A high level of correlation (92.6%) between morphological and molecular identifications was found.

Development and application of two independent real-time PCR assays to detect clinically relevant Mucorales species

“…Using 10-fold dilutions of genomic DNA, standard curves were generated. Screening of recent publications on Mucorales infections (Alvarez et al, 2009;Rüping et al, 2010;Skiada et al, 2011;Lanternier et al, 2012;Roden et al, 2005) led to the identification of the three most common Mucorales species in each manuscript. In total, we found five different Mucorales species, which were further investigated by serial dilution.…”

Identification of fungal pathogens in a patient with acute myelogenic leukemia using a pathogen detection array technology

“…4 Thus, molecular assays were developed to identify specific species of zygomycetes. These include sequencing of internal transcribed spacer (ITS) region, [5][6][7][8] 28S rRNA 9 and non-sequencing based tests like polymerase chain reaction (PCR) with restriction fragment length polymorphism (RFLP), 10,11 real-time PCR using melting curve analysis, 12 multiplex PCR using specific DNA probes in a microarray based assay, 13 and PCR assay using the Luminex xMAP hybridization method.…”

Adaptation to thermotolerance inRhizopuscoincides with virulence as revealed by avian and invertebrate infection models, phylogeny, physiological and metabolic flexibility

“…16 While R. arrhizus and R. stolonifer are found to similar extends, the majority of infections is caused by R. arrhizus (50%) and R. microsporus (15-25%). 16,17,43 This could be explained by the lower virulence potential of R. stolonifer observed in this study. In contrast, R. schipperae, R. caespitosus and R. homothallicus appear to be less abundant in the environment, if judging from the few available specimens from public culture collections, or the fact, that R. schipperae is only known from 2 reported cases with no obvious natural substrate presented.…”