Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis
Aspergillus fumigatus triazole resistance is an emerging concern for treating chronically infected/colonized patients. This study sought to evaluate the performance of PCR assays to detect Aspergillus fungi together with azole resistance in sputum samples from cystic fibrosis (CF) patients. In total...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-03-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fmicb.2018.00531/full |
| _version_ | 1818941026935504896 |
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| author | Hélène Guegan Sylviane Chevrier Chantal Belleguic Eric Deneuville Florence Robert-Gangneux Florence Robert-Gangneux Jean-Pierre Gangneux Jean-Pierre Gangneux |
| author_facet | Hélène Guegan Sylviane Chevrier Chantal Belleguic Eric Deneuville Florence Robert-Gangneux Florence Robert-Gangneux Jean-Pierre Gangneux Jean-Pierre Gangneux |
| author_sort | Hélène Guegan |
| collection | DOAJ |
| description | Aspergillus fumigatus triazole resistance is an emerging concern for treating chronically infected/colonized patients. This study sought to evaluate the performance of PCR assays to detect Aspergillus fungi together with azole resistance in sputum samples from cystic fibrosis (CF) patients. In total, 119 sputum samples from 87 CF patients were prospectively processed for Aspergillus detection by means of mycological culture and four qPCR assays, 2 in-house methods and two commercial multiplex real-time PCR assays simultaneously detecting Aspergillus and the most relevant cyp51A gene mutations (MycoGENIE® and AsperGenius®). Azole susceptibility of A. fumigatus isolates was assessed using Etest® method and cyp51A gene mutation were characterized by sequencing. The overall rate of Aspergillus detection with the four qPCR assays ranged from 47.9 to 57.1%, contrasting with 42/119 (35.3%) positive cultures with A. fumigatus. The high sensitivity of PCR on sputum could then contribute to more effective grading of Aspergillus disease in CF patients. Five out of 41 isolated strains (12.2%) exhibited azole-resistant MIC patterns, three of which harbored cyp51A mutations and only 1/3 with the sequence TR34/L98H. Combined with culture, PCR assay achieved high sensitivity Aspergillus screening in CF samples. However, cyp51A targeting was only moderately effective for azole resistance monitoring, while Aspergillus resistance remains of great concern. |
| first_indexed | 2024-12-20T06:49:00Z |
| format | Article |
| id | doaj.art-0d5c1614262e44e3812ea467da430280 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-20T06:49:00Z |
| publishDate | 2018-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-0d5c1614262e44e3812ea467da4302802022-12-21T19:49:37ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-03-01910.3389/fmicb.2018.00531345974Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic FibrosisHélène Guegan0Sylviane Chevrier1Chantal Belleguic2Eric Deneuville3Florence Robert-Gangneux4Florence Robert-Gangneux5Jean-Pierre Gangneux6Jean-Pierre Gangneux7Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, Rennes, FranceLaboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, Rennes, FranceCentre de Ressources et de Compétences de la Mucoviscidose Adulte, Centre Hospitalier Universitaire de Rennes, Rennes, FranceCentre de Ressources et de Compétences de la Mucoviscidose Pédiatrique, Centre Hospitalier Universitaire de Rennes, Rennes, FranceLaboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, Rennes, FranceUniversité de Rennes 1, INSERM, Institut de Recherche en Santé, Environnement et Travail – UMR_S 1085, Rennes, FranceLaboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, Rennes, FranceUniversité de Rennes 1, INSERM, Institut de Recherche en Santé, Environnement et Travail – UMR_S 1085, Rennes, FranceAspergillus fumigatus triazole resistance is an emerging concern for treating chronically infected/colonized patients. This study sought to evaluate the performance of PCR assays to detect Aspergillus fungi together with azole resistance in sputum samples from cystic fibrosis (CF) patients. In total, 119 sputum samples from 87 CF patients were prospectively processed for Aspergillus detection by means of mycological culture and four qPCR assays, 2 in-house methods and two commercial multiplex real-time PCR assays simultaneously detecting Aspergillus and the most relevant cyp51A gene mutations (MycoGENIE® and AsperGenius®). Azole susceptibility of A. fumigatus isolates was assessed using Etest® method and cyp51A gene mutation were characterized by sequencing. The overall rate of Aspergillus detection with the four qPCR assays ranged from 47.9 to 57.1%, contrasting with 42/119 (35.3%) positive cultures with A. fumigatus. The high sensitivity of PCR on sputum could then contribute to more effective grading of Aspergillus disease in CF patients. Five out of 41 isolated strains (12.2%) exhibited azole-resistant MIC patterns, three of which harbored cyp51A mutations and only 1/3 with the sequence TR34/L98H. Combined with culture, PCR assay achieved high sensitivity Aspergillus screening in CF samples. However, cyp51A targeting was only moderately effective for azole resistance monitoring, while Aspergillus resistance remains of great concern.http://journal.frontiersin.org/article/10.3389/fmicb.2018.00531/fullAspergillusaspergillosiscystic fibrosisquantitative real-time PCRazole resistancesputum |
| spellingShingle | Hélène Guegan Sylviane Chevrier Chantal Belleguic Eric Deneuville Florence Robert-Gangneux Florence Robert-Gangneux Jean-Pierre Gangneux Jean-Pierre Gangneux Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis Frontiers in Microbiology Aspergillus aspergillosis cystic fibrosis quantitative real-time PCR azole resistance sputum |
| title | Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis |
| title_full | Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis |
| title_fullStr | Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis |
| title_full_unstemmed | Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis |
| title_short | Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis |
| title_sort | performance of molecular approaches for aspergillus detection and azole resistance surveillance in cystic fibrosis |
| topic | Aspergillus aspergillosis cystic fibrosis quantitative real-time PCR azole resistance sputum |
| url | http://journal.frontiersin.org/article/10.3389/fmicb.2018.00531/full |
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