Abstract
Diagnosis of invasive fungal infections is complex, and the lack of standardization of molecular methods is still a challenge. Several methods are available for the diagnosis of invasive aspergillosis, but their effectiveness will depend on the studied population, the patients’ comorbidities, and the use of mold active prophylaxis, among others. The ability to determine the identity of the infecting Aspergillus species, and to detect mutations conferring specific resistance patterns directly from DNA extracted from the biological product, is an advantage of nucleic acid testing compared with antigen-based assays. In this study, we present laboratory cases where the diagnosis of aspergillosis was performed using a real-time multiplex PCR for the detection of Aspergillus DNA in tissue samples, showing its usefulness as one more tool in the diagnosis of aspergillosis in tissue samples. Aspergillus real-time multiplex PCR was also used to detect azole-resistance in some cases. In the majority of the PCR positive cases, cultures remained negative after 60 days. The PCR assay directed to Aspergillus gave positive signals for Aspergillus fumigatus sensu stricto. Results were confirmed by panfungal PCR, followed by sequencing, revealing 100% homology with Aspergillus fumigatus sensu stricto. Mutations conferring azole resistance were not detected.
Highlights
Invasive fungal infections have significantly increased due to advances in medical care in the at-risk immunocompromised population [1]
PCR based detection and identification of Aspergillus in tissue samples, together with the detection of possible mutations that confer azole resistance to azoles, enable the mycology laboratory to give a prompt result to clinicians, in order to speed up diagnosis for early appropriate treatment
The aim of this study is to present laboratory cases of possible/proven invasive aspergillosis where this commercially available kit was applied and show that despite not being validated to tissue samples yet, this method may help in the diagnosis and decisions on therapy regimens
Summary
Invasive fungal infections have significantly increased due to advances in medical care in the at-risk immunocompromised population [1]. Changes in the spectrum of the fungal infections associated with new risk factors and the emergence of resistant fungi highlight the need for continuous updates on knowledge of the epidemiology of fungal infections [2]. Aspergillus is the filamentous fungi more frequently associated with invasive fungal infection. According to the LIFE organization, more than 30 million patients are at risk for invasive aspergillosis and about 300,000 patients will develop the disease annually [3]. Patients at higher risk of acquiring invasive aspergillosis (IA) often suffer from severe granulocytopenia (i.e., leukemia and bone marrow or solid organ transplanted patients). Long-term corticosteroid use, diabetes, major burns, and recent major surgeries are considered as predisposing risk factors. Diagnosis of invasive aspergillosis (IA) is a challenge and should be based on the integration of clinical, radiological, and microbiological data
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