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Molecular Detection of Aspergillus: 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’ comorbidities, and th...

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Autores principales: Sabino, Raquel, Simões, Helena, Veríssimo, Cristina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7151104/
https://www.ncbi.nlm.nih.gov/pubmed/31936735
http://dx.doi.org/10.3390/jof6010011
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author Sabino, Raquel
Simões, Helena
Veríssimo, Cristina
author_facet Sabino, Raquel
Simões, Helena
Veríssimo, Cristina
author_sort Sabino, Raquel
collection PubMed
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 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.
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spelling pubmed-71511042020-04-20 Molecular Detection of Aspergillus: Application of a Real-Time PCR Multiplex Assay in Tissue Samples Sabino, Raquel Simões, Helena Veríssimo, Cristina J Fungi (Basel) Brief Report 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. MDPI 2020-01-10 /pmc/articles/PMC7151104/ /pubmed/31936735 http://dx.doi.org/10.3390/jof6010011 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Brief Report
Sabino, Raquel
Simões, Helena
Veríssimo, Cristina
Molecular Detection of Aspergillus: Application of a Real-Time PCR Multiplex Assay in Tissue Samples
title Molecular Detection of Aspergillus: Application of a Real-Time PCR Multiplex Assay in Tissue Samples
title_full Molecular Detection of Aspergillus: Application of a Real-Time PCR Multiplex Assay in Tissue Samples
title_fullStr Molecular Detection of Aspergillus: Application of a Real-Time PCR Multiplex Assay in Tissue Samples
title_full_unstemmed Molecular Detection of Aspergillus: Application of a Real-Time PCR Multiplex Assay in Tissue Samples
title_short Molecular Detection of Aspergillus: Application of a Real-Time PCR Multiplex Assay in Tissue Samples
title_sort molecular detection of aspergillus: application of a real-time pcr multiplex assay in tissue samples
topic Brief Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7151104/
https://www.ncbi.nlm.nih.gov/pubmed/31936735
http://dx.doi.org/10.3390/jof6010011
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