Direct molecular diagnosis of aspergillosis and CYP51A profiling from respiratory samples of French patients
Background: Microbiological diagnosis of aspergillosis and triazole resistance is limited by poor culture yield. To better estimate this shortcoming, we compared culture and molecular detection of A. fumigatus in respiratory samples from French patients at risk for aspergillosis. Methods: A total of...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2016-07-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.01164/full |
| _version_ | 1811326452685078528 |
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| author | Yanan Zhao Cécile Garnaud Cécile Garnaud Marie-Pierre Brenier-Pinchart Marie-Pierre Brenier-Pinchart Anne Thiébaut-Bertrand Anne Thiébaut-Bertrand Christel Saint-Raymond Boubou Camara Rebecca Hamidfar-Roy Odile Cognet Danièle Maubon Danièle Maubon Muriel Cornet Muriel Cornet David S Perlin |
| author_facet | Yanan Zhao Cécile Garnaud Cécile Garnaud Marie-Pierre Brenier-Pinchart Marie-Pierre Brenier-Pinchart Anne Thiébaut-Bertrand Anne Thiébaut-Bertrand Christel Saint-Raymond Boubou Camara Rebecca Hamidfar-Roy Odile Cognet Danièle Maubon Danièle Maubon Muriel Cornet Muriel Cornet David S Perlin |
| author_sort | Yanan Zhao |
| collection | DOAJ |
| description | Background: Microbiological diagnosis of aspergillosis and triazole resistance is limited by poor culture yield. To better estimate this shortcoming, we compared culture and molecular detection of A. fumigatus in respiratory samples from French patients at risk for aspergillosis. Methods: A total of 97 respiratory samples including bronchoalveolar lavages (BAL), bronchial aspirates (BA), tracheal aspirates, sputa, pleural fluids, and lung biopsy were collected from 33 patients having invasive aspergillosis (n=12), chronic pulmonary aspergillosis (n=3), allergic bronchopulmonary aspergillosis (n=7) or colonization (n=11) and 28 controls. Each specimen was evaluated by culture, pan-Aspergillus qPCR, and CYP51A PCR and sequencing. Results: One A. flavus and 19 A. fumigatus with one multiazole resistant strain (5.3%) were cultured from 20 samples. Culture positivity was 62.5%, 75%, 42.9%, and 15.8% in ABPA, CPA, IA and colonized patients, respectively. Aspergillus detection rate was significantly higher by pan-Aspergillus qPCR than by culture in IA (90.5% vs 42.9%; P<0.05) and colonization group (73.7% vs 15.8%; P<0.05). The CYP51A PCR found one TR34/L98H along with 5 novel cyp51A mutations (4 non-synonymous and 1 promoter mutations), yet no association can be established currently between these novel mutations and azole resistance. The analysis of 11 matched pairs of BA and BAL samples found that 9/11 BA carried greater fungal load than BAL and CYP51A detection was more sensitive in BA than in BAL. Conclusion: Direct molecular detection of Aspergillus spp. and azole resistance markers are useful adjunct tools for comprehensive aspergillosis diagnosis. The observed superior diagnostic value of BAs to BAL fluids warrants more in-depth study. |
| first_indexed | 2024-04-13T14:50:33Z |
| format | Article |
| id | doaj.art-28bb9c98204940cfa3e0be09591bfd8d |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-13T14:50:33Z |
| publishDate | 2016-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-28bb9c98204940cfa3e0be09591bfd8d2022-12-22T02:42:37ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2016-07-01710.3389/fmicb.2016.01164205495Direct molecular diagnosis of aspergillosis and CYP51A profiling from respiratory samples of French patientsYanan Zhao0Cécile Garnaud1Cécile Garnaud2Marie-Pierre Brenier-Pinchart3Marie-Pierre Brenier-Pinchart4Anne Thiébaut-Bertrand5Anne Thiébaut-Bertrand6Christel Saint-Raymond7Boubou Camara8Rebecca Hamidfar-Roy9Odile Cognet10Danièle Maubon11Danièle Maubon12Muriel Cornet13Muriel Cornet14David S Perlin15Public Health Research Institute, New Jersey Medical School, Rutgers Biomedical and Health SciencesLaboratoire de Parasitologie-Mycologie, Institut de Biologie et Pathologie, Centre Hospitalier Universitaire Grenoble AlpesLaboratoire TIMC-IMAG-TheREx, UMR 5525 CNRS, Université Grenoble AlpesLaboratoire de Parasitologie-Mycologie, Institut de Biologie et Pathologie, Centre Hospitalier Universitaire Grenoble AlpesInstitut Albert Bonniot, CNRS UMR 5309, INSERM U1209, Université Grenoble-AlpesLaboratoire TIMC-IMAG-TheREx, UMR 5525 CNRS, Université Grenoble AlpesClinique Universitaire d’Hématologie, Centre Hospitalier Universitaire Grenoble AlpesClinique Universitaire de Pneumologie, Centre Hospitalier Universitaire Grenoble AlpesClinique Universitaire de Pneumologie, Centre Hospitalier Universitaire Grenoble AlpesRéanimation Médicale, Centre Hospitalier Universitaire Grenoble AlpesLaboratoire de Parasitologie-Mycologie, Institut de Biologie et Pathologie, Centre Hospitalier Universitaire Grenoble AlpesLaboratoire de Parasitologie-Mycologie, Institut de Biologie et Pathologie, Centre Hospitalier Universitaire Grenoble AlpesLaboratoire TIMC-IMAG-TheREx, UMR 5525 CNRS, Université Grenoble AlpesLaboratoire de Parasitologie-Mycologie, Institut de Biologie et Pathologie, Centre Hospitalier Universitaire Grenoble AlpesLaboratoire TIMC-IMAG-TheREx, UMR 5525 CNRS, Université Grenoble AlpesPublic Health Research Institute, New Jersey Medical School, Rutgers Biomedical and Health SciencesBackground: Microbiological diagnosis of aspergillosis and triazole resistance is limited by poor culture yield. To better estimate this shortcoming, we compared culture and molecular detection of A. fumigatus in respiratory samples from French patients at risk for aspergillosis. Methods: A total of 97 respiratory samples including bronchoalveolar lavages (BAL), bronchial aspirates (BA), tracheal aspirates, sputa, pleural fluids, and lung biopsy were collected from 33 patients having invasive aspergillosis (n=12), chronic pulmonary aspergillosis (n=3), allergic bronchopulmonary aspergillosis (n=7) or colonization (n=11) and 28 controls. Each specimen was evaluated by culture, pan-Aspergillus qPCR, and CYP51A PCR and sequencing. Results: One A. flavus and 19 A. fumigatus with one multiazole resistant strain (5.3%) were cultured from 20 samples. Culture positivity was 62.5%, 75%, 42.9%, and 15.8% in ABPA, CPA, IA and colonized patients, respectively. Aspergillus detection rate was significantly higher by pan-Aspergillus qPCR than by culture in IA (90.5% vs 42.9%; P<0.05) and colonization group (73.7% vs 15.8%; P<0.05). The CYP51A PCR found one TR34/L98H along with 5 novel cyp51A mutations (4 non-synonymous and 1 promoter mutations), yet no association can be established currently between these novel mutations and azole resistance. The analysis of 11 matched pairs of BA and BAL samples found that 9/11 BA carried greater fungal load than BAL and CYP51A detection was more sensitive in BA than in BAL. Conclusion: Direct molecular detection of Aspergillus spp. and azole resistance markers are useful adjunct tools for comprehensive aspergillosis diagnosis. The observed superior diagnostic value of BAs to BAL fluids warrants more in-depth study.http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.01164/fullAspergillus fumigatusMolecular diagnosticsAzole resistanceRespiratory samplesCYP51AFrench patients |
| spellingShingle | Yanan Zhao Cécile Garnaud Cécile Garnaud Marie-Pierre Brenier-Pinchart Marie-Pierre Brenier-Pinchart Anne Thiébaut-Bertrand Anne Thiébaut-Bertrand Christel Saint-Raymond Boubou Camara Rebecca Hamidfar-Roy Odile Cognet Danièle Maubon Danièle Maubon Muriel Cornet Muriel Cornet David S Perlin Direct molecular diagnosis of aspergillosis and CYP51A profiling from respiratory samples of French patients Frontiers in Microbiology Aspergillus fumigatus Molecular diagnostics Azole resistance Respiratory samples CYP51A French patients |
| title | Direct molecular diagnosis of aspergillosis and CYP51A profiling from respiratory samples of French patients |
| title_full | Direct molecular diagnosis of aspergillosis and CYP51A profiling from respiratory samples of French patients |
| title_fullStr | Direct molecular diagnosis of aspergillosis and CYP51A profiling from respiratory samples of French patients |
| title_full_unstemmed | Direct molecular diagnosis of aspergillosis and CYP51A profiling from respiratory samples of French patients |
| title_short | Direct molecular diagnosis of aspergillosis and CYP51A profiling from respiratory samples of French patients |
| title_sort | direct molecular diagnosis of aspergillosis and cyp51a profiling from respiratory samples of french patients |
| topic | Aspergillus fumigatus Molecular diagnostics Azole resistance Respiratory samples CYP51A French patients |
| url | http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.01164/full |
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