key: cord-0252235-tv9hxbgr authors: Shirvani, Fariba; Fattahi, Azam title: Pulmonary Candidiasis Associated with COVID-19: Evaluation of Causative Agents and their Antifungal Susceptibility Patterns date: 2021-01-03 journal: Tanaffos DOI: nan sha: 043c96df456a6068704baa4f2b6ff109543e269f doc_id: 252235 cord_uid: tv9hxbgr BACKGROUND: The purpose of the present study was to isolate Candida species from individuals with the COVID-19 disease and evaluate the susceptibility pattern of Candida spp. to routine antifungal drugs. MATERIALS AND METHODS: A total of 25 Candida spp. isolated from hospitalized patients with COVID-19, who were suspected to have pulmonary candidiasis, and 26 archived Candida spp. specimens were enrolled in this study. For the identification of Candida spp., PCR was performed to detect and amplify the ITS1 and ITS4 genes. Then the products were subjected to the Msp I restriction enzyme to precisely identify the species. The amplification of the WHP1 gene was conducted to identify Candida albicans species. The antifungal activities of routine drugs and the synthesize AuNPs against Candida spp. were assessed based on the protocols presented by the Clinical and Laboratory Standards Institute M60. RESULTS: In the present study, C. albicans (24; 96%) and C. parapsilosis (1; 4%) were identified as the etiologic agents of the pulmonary candidiasis associated with the COVID-19 infection. Voriconazol and amphotericin B had superior activity against all the isolates in this study. Treatment with fluconazole and itraconazole did not significantly change the formation of colony-forming units (CFU). However, treatment with the AuNPs significantly decreased (within the range of 92–99.1%; P<0.05) the number of CFUs. CONCLUSION: The azole prophylaxis has likely been associated with the development of resistant isolates; the results of the present study suggested the promising role of novel antifungal agents such as AuNPs in overcoming drug resistant fungi. Nanoparticles (NPs) have supposedly higher in vitro activities against C. albicans than more recently developed and probably more active azoles (10, 11) . They are also among valuable options for drug delivery. The combination of NPs with antifungals decreases the risk of toxicity and increases the activities of antifungals (12) . Genomic DNA was directly isolated from BAL specimens using a high-pure PCR templet purification kit (Roche, Germany) according to the manufacturer's user guide. The extracted DNA was stored at -20ºC until further analysis. To identify Candida spp., PCR was performed to detect the ITS1 and ITS4 genes using the following protocol: gel. The amplification of the WHP1 gene was conducted to identify C. albicans, according to a previous report (13) . For the characterization of the sizes and charges of the AuNPs, the Z average size and Zeta potential were estimated using the Malvern Zetasizer Nano ZS90 instrument (Malvern Panalytical Ltd., England). The Membrane permeability changes in the C. albicans and C. parapsilosis exposed to AuNPs were estimated via Statistical analysis was performed using SPSS software (version 16.0). The non-parametric Mann-Whitney U test was utilized to compare the results. According to molecular analyses, only C. albicans (24; 96%) and C. parapsilosis (1; 4%) ( Figure 1A A recently developed delivery system for antifungals can be used to enhance the fungicidal properties of AuNPs (25, 26) . The findings of the present study raised the hypothesis that the fungicidal activity of azoles against resistant fungi can be enhanced in conjugation with AuNPs. 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