Molecular Detection of <i>Aspergillus</i>: Application of a Real-Time PCR Multiplex Assay in Tissue Samples
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’ comorbiditi...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-01-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/6/1/11 |
| _version_ | 1819010278030835712 |
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| author | Raquel Sabino Helena Simões Cristina Veríssimo |
| author_facet | Raquel Sabino Helena Simões Cristina Veríssimo |
| author_sort | Raquel Sabino |
| collection | DOAJ |
| description | 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 <i>Aspergillus</i> 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 <i>Aspergillus</i> DNA in tissue samples, showing its usefulness as one more tool in the diagnosis of aspergillosis in tissue samples. <i>Aspergillus</i> 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 <i>Aspergillus</i> gave positive signals for <i>Aspergillus fumigatus</i> sensu stricto. Results were confirmed by panfungal PCR, followed by sequencing, revealing 100% homology with <i>Aspergillus fumigatus</i> sensu stricto. Mutations conferring azole resistance were not detected. |
| first_indexed | 2024-12-21T01:09:43Z |
| format | Article |
| id | doaj.art-476aa53f07ff4adaab09f7122eccf51d |
| institution | Directory Open Access Journal |
| issn | 2309-608X |
| language | English |
| last_indexed | 2024-12-21T01:09:43Z |
| publishDate | 2020-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj.art-476aa53f07ff4adaab09f7122eccf51d2022-12-21T19:20:57ZengMDPI AGJournal of Fungi2309-608X2020-01-01611110.3390/jof6010011jof6010011Molecular Detection of <i>Aspergillus</i>: Application of a Real-Time PCR Multiplex Assay in Tissue SamplesRaquel Sabino0Helena Simões1Cristina Veríssimo2Reference Unit for Parasitic and Fungal Infections, Department of Infectious Diseases, National Institute of Health Dr. Ricardo Jorge, Av. Padre Cruz, 1649-016 Lisbon, PortugalReference Unit for Parasitic and Fungal Infections, Department of Infectious Diseases, National Institute of Health Dr. Ricardo Jorge, Av. Padre Cruz, 1649-016 Lisbon, PortugalReference Unit for Parasitic and Fungal Infections, Department of Infectious Diseases, National Institute of Health Dr. Ricardo Jorge, Av. Padre Cruz, 1649-016 Lisbon, PortugalDiagnosis 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 <i>Aspergillus</i> 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 <i>Aspergillus</i> DNA in tissue samples, showing its usefulness as one more tool in the diagnosis of aspergillosis in tissue samples. <i>Aspergillus</i> 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 <i>Aspergillus</i> gave positive signals for <i>Aspergillus fumigatus</i> sensu stricto. Results were confirmed by panfungal PCR, followed by sequencing, revealing 100% homology with <i>Aspergillus fumigatus</i> sensu stricto. Mutations conferring azole resistance were not detected.https://www.mdpi.com/2309-608X/6/1/11<i>aspergillus</i>invasive fungal infectionsreal-time pcrmolecular diagnosisazole-resistance |
| spellingShingle | Raquel Sabino Helena Simões Cristina Veríssimo Molecular Detection of <i>Aspergillus</i>: Application of a Real-Time PCR Multiplex Assay in Tissue Samples Journal of Fungi <i>aspergillus</i> invasive fungal infections real-time pcr molecular diagnosis azole-resistance |
| title | Molecular Detection of <i>Aspergillus</i>: Application of a Real-Time PCR Multiplex Assay in Tissue Samples |
| title_full | Molecular Detection of <i>Aspergillus</i>: Application of a Real-Time PCR Multiplex Assay in Tissue Samples |
| title_fullStr | Molecular Detection of <i>Aspergillus</i>: Application of a Real-Time PCR Multiplex Assay in Tissue Samples |
| title_full_unstemmed | Molecular Detection of <i>Aspergillus</i>: Application of a Real-Time PCR Multiplex Assay in Tissue Samples |
| title_short | Molecular Detection of <i>Aspergillus</i>: Application of a Real-Time PCR Multiplex Assay in Tissue Samples |
| title_sort | molecular detection of i aspergillus i application of a real time pcr multiplex assay in tissue samples |
| topic | <i>aspergillus</i> invasive fungal infections real-time pcr molecular diagnosis azole-resistance |
| url | https://www.mdpi.com/2309-608X/6/1/11 |
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