{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:28:39Z","timestamp":1760239719564,"version":"build-2065373602"},"reference-count":49,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2020,12,9]],"date-time":"2020-12-09T00:00:00Z","timestamp":1607472000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Aspergillus fumigatus is the main causative agent of Invasive Aspergillosis. This mold produces conidia that when inhaled by immunocompromized hosts can be deposited in the lungs and germinate, triggering disease. In this paper, the development of a method using peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) is described. The PNA-FISH probe was tested in several strains and a specificity and sensitivity of 100% was obtained. Detection of A. fumigatussensu stricto was then achieved in artificial sputum medium (ASM) pre-inoculated with 1 \u00d7 102 conidia\u00b7mL\u22121\u20131 \u00d7 103 conidia\u00b7mL\u22121, after a germination step of 24 h. The PNA-FISH method was evaluated in 24 clinical samples (10 sputum, 8 bronchoalveolar lavage (BAL), and 6 bronchial lavage (BL)) that were inoculated with 1 \u00d7 104 conidia\u00b7mL\u22121 in sputum; 1 \u00d7 103 conidia\u00b7mL\u22121 in BL and BAL, for 24 h. Despite a specificity of 100%, the sensitivity was 79%. This relatively low sensitivity can be explained by the fact that hyphae can yield \u201cfungal ball\u201c clusters, hindering pipetting procedures and subsequent detection, leading to false negative results. Nonetheless, this study showed the potential of the PNA-FISH method for A. fumigatussensu stricto detection since it takes only 1 h 30 m to perform the procedure with a pre-enrichment step of 6 h (pure cultures) and 24 h (clinical samples), and might provide a suitable alternative to the existing methods for studies in pure cultures and in clinical settings.<\/jats:p>","DOI":"10.3390\/microorganisms8121950","type":"journal-article","created":{"date-parts":[[2020,12,9]],"date-time":"2020-12-09T09:17:58Z","timestamp":1607505478000},"page":"1950","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Establishment of a New PNA-FISH Method for Aspergillus fumigatus Identification: First Insights for Future Use in Pulmonary Samples"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8452-4749","authenticated-orcid":false,"given":"Laura","family":"Cerqueira","sequence":"first","affiliation":[{"name":"LEPABE\u2014Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal"},{"name":"BIOMODE 2, S.A., Edif\u00edcio GNRATION, Pra\u00e7a Conde Agrolongo n\u00ba 123, 4700-312 Braga, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5791-6528","authenticated-orcid":false,"given":"Sara","family":"Moura","sequence":"additional","affiliation":[{"name":"BIOMODE 2, S.A., Edif\u00edcio GNRATION, Pra\u00e7a Conde Agrolongo n\u00ba 123, 4700-312 Braga, Portugal"},{"name":"Laboratory of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Coimbra, 3000-548 Coimbra, Portugal"},{"name":"Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0175-1119","authenticated-orcid":false,"given":"Carina","family":"Almeida","sequence":"additional","affiliation":[{"name":"BIOMODE 2, S.A., Edif\u00edcio GNRATION, Pra\u00e7a Conde Agrolongo n\u00ba 123, 4700-312 Braga, Portugal"},{"name":"INIAV, IP\u2014National Institute for Agrarian and Veterinary Research, Rua dos Lagidos, Lugar da Madalena, 4485-655 Vair\u00e3o, Vila do Conde, Portugal"}]},{"given":"Maria Jo\u00e3o","family":"Vieira","sequence":"additional","affiliation":[{"name":"CEB\u2014Centre of Biological Engineering, LIBRO\u2014Laboratory of Research in Biofilms Ros\u00e1rio Oliveira, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal"}]},{"given":"Nuno Filipe","family":"Azevedo","sequence":"additional","affiliation":[{"name":"LEPABE\u2014Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2020,12,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1099\/mic.0.000651","article-title":"Microbe Profile: Aspergillus fumigatus: A saprotrophic and opportunistic fungal pathogen","volume":"164","author":"Fang","year":"2018","journal-title":"Microbiology (Reading)"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e1003743","DOI":"10.1371\/journal.ppat.1003743","article-title":"Aspergillus fumigatus\u2014what makes the species a ubiquitous human fungal pathogen?","volume":"9","author":"Sugui","year":"2013","journal-title":"PLoS Pathog."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.riam.2010.10.003","article-title":"What makes Aspergillus fumigatus a successful pathogen? 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