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PCR based identification and discrimination of agents of mucormycosis and aspergillosis in paraffin wax embedded tissue

¹ Institute for Tropical Medicine, University Hospital Tübingen, Keplerstrasse 15, 72074 Tübingen, Germany
² Institute for Medical Microbiology, Virology and Hygiene, University Hospital Rostock, Rostock D-18057, Germany
³ Departamento de Pathologia, Universidad Autonoma Nuevo Leon, Monterrey, NL, Mexico CP 64460
⁴ Departamento de Microbiologia, Universidad Autonoma Nuevo
⁵ Medizinische Klinik III, University Hospital, D-60590 Frankfurt/Main, Germany
⁶ Monterrey, Mexico, and Institute of Hygiene and Medical Microbiology, Medical University of Vienna, A-1090 Wein, Austria
⁷ Department of Medicine, Division of Infectious Diseases and Hospital Epidemiology, CH-8091 Zurich, Switzerland
⁸ Department of Hygiene and Social Medicine, University of Innsbruck, A-6010 Innsbruck, Austria

Correspondence to:
Dr R Bialek
Institut für Tropenmedizin, Universitätsklinikum Tübingen, Keplerstrasse 15, 72074 Tübingen, Germany; ralf.bialek{at}med.uni-tuebingen.de Background: Invasive fungal infections are often diagnosed by histopathology without identification of the causative fungi, which show significantly different antifungal susceptibilities.

Aims: To establish and evaluate a system of two seminested polymerase chain reaction (PCR) assays to identify and discriminate between agents of aspergillosis and mucormycosis in paraffin wax embedded tissue samples.

Methods: DNA of 52 blinded samples from five different centres was extracted and used as a template in two PCR assays targeting the mitochondrial aspergillosis DNA and the 18S ribosomal DNA of zygomycetes.

Results: Specific fungal DNA was identified in 27 of 44 samples in accordance with a histopathological diagnosis of zygomycosis or aspergillosis, respectively. Aspergillus fumigatus DNA was amplified from one specimen of zygomycosis (diagnosed by histopathology). In four of 16 PCR negative samples no human DNA was amplified, possibly as a result of the destruction of DNA before paraffin wax embedding. In addition, eight samples from clinically suspected fungal infections (without histopathological proof) were examined. The two PCR assays detected a concomitant infection with Absidia corymbifera and A fumigatus in one, and infections with Rhizopus arrhizus and A fumigatus in another two cases.

Conclusions: The two seminested PCR assays described here can support a histopathological diagnosis of mucormycosis or aspergillosis, and can identify the infective agent, thereby optimising antifungal treatment.

Abbreviations: PCR, polymerase chain reaction; rDNA, ribosomal DNA

Keywords: mucormycosis; aspergillosis; zygomycetes; polymerase chain reaction