title: Direct Molecular Diagnosis of Aspergillosis and CYP51A Profiling from Respiratory Samples of French Patients.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: New Jersey Medical School, Public Health Research Institute, Rutgers Biomedical and Health Sciences Newark, NJ, USA.
authors.forename: Yanan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Institut de Biologie et Pathologie, Centre Hospitalier Universitaire Grenoble AlpesGrenoble, France; Laboratoire TIMC-IMAG-TheREx, UMR 5525 Centre National de la Recherche Scientifique, Université Grenoble AlpesGrenoble, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Garnaud
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Institut de Biologie et Pathologie, Centre Hospitalier Universitaire Grenoble AlpesGrenoble, France; Institut Albert Bonniot, Centre National de la Recherche Scientifique UMR 5309, Institut National de la Santé et de la Recherche Médicale U1209, Université Grenoble-AlpesGrenoble, France.
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Brenier-Pinchart
authors.affiliation: Laboratoire TIMC-IMAG-TheREx, UMR 5525 Centre National de la Recherche Scientifique, Université Grenoble AlpesGrenoble, France; Clinique Universitaire d'Hématologie, Centre Hospitalier Universitaire Grenoble AlpesGrenoble, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Thiébaut-Bertrand
authors.affiliation: Clinique Universitaire de Pneumologie, Centre Hospitalier Universitaire Grenoble Alpes Grenoble, France.
authors.forename: Christel
authors.initials: C
authors.lastname: Saint-Raymond
authors.affiliation: Clinique Universitaire de Pneumologie, Centre Hospitalier Universitaire Grenoble Alpes Grenoble, France.
authors.forename: Boubou
authors.initials: B
authors.lastname: Camara
authors.affiliation: Réanimation Médicale, Centre Hospitalier Universitaire Grenoble Alpes Grenoble, France.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Hamidfar
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Institut de Biologie et Pathologie, Centre Hospitalier Universitaire Grenoble Alpes Grenoble, France.
authors.forename: Odile
authors.initials: O
authors.lastname: Cognet
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Institut de Biologie et Pathologie, Centre Hospitalier Universitaire Grenoble AlpesGrenoble, France; Laboratoire TIMC-IMAG-TheREx, UMR 5525 Centre National de la Recherche Scientifique, Université Grenoble AlpesGrenoble, France.
authors.forename: Danièle
authors.initials: D
authors.lastname: Maubon
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Institut de Biologie et Pathologie, Centre Hospitalier Universitaire Grenoble AlpesGrenoble, France; Laboratoire TIMC-IMAG-TheREx, UMR 5525 Centre National de la Recherche Scientifique, Université Grenoble AlpesGrenoble, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Cornet
authors.affiliation: New Jersey Medical School, Public Health Research Institute, Rutgers Biomedical and Health Sciences Newark, NJ, USA.
authors.forename: David S
authors.initials: DS
authors.lastname: Perlin
keywords: Aspergillus fumigatus; CYP51A; French patients; azole resistance; molecular diagnostics; respiratory samples