Hélène Guegan1, Florence Robert-Gangneux2, Christophe Camus3, Sorya Belaz2, Tony Marchand4, Marion Baldeyrou5, Jean-Pierre Gangneux6. 1. Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, Rennes, France. 2. Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, Rennes, France; Univ Rennes, Inserm, EHESP, Irset (Institut de Recherche en santé, environnement et travail) - UMR_S 1085, Rennes, France. 3. Service de réanimation médicale, Centre Hospitalier Universitaire de Rennes, Rennes, France. 4. Service d'hématologie clinique, Centre Hospitalier Universitaire de Rennes, Rennes, France. 5. Service de Maladies Infectieuses, Centre Hospitalier Universitaire de Rennes, Rennes, France. 6. Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Rennes, Rennes, France; Univ Rennes, Inserm, EHESP, Irset (Institut de Recherche en santé, environnement et travail) - UMR_S 1085, Rennes, France. Electronic address: jean-pierre.gangneux@univ-rennes1.fr.
Abstract
OBJECTIVES: This study aimed to evaluate new tools to diagnose invasive aspergillosis (IA) directly from broncho-alveolar lavage (BAL) samples. METHODS: All consecutive patients with suspected IA who underwent bronchoscopy with BAL were prospectively included. Mycological culture and ELISA detection of galactomannan (GM) were performed on BAL. Two in-house and two marketed PCR assays were used on BAL DNA extracts to detect Aspergillus species. Susceptibility testing was performed after culture; marketed PCR assays detected mutations in the CYP51A gene associated to resistance. RESULTS: Within 3 years, 1555 BAL samples were processed, including 413 samples from 387 immunosuppressed patients. IA diagnosis was no-IA, possible, probable or proven IA in 326, 23, 37 and 1 patients, respectively. PCR assays sensitivity for Aspergillus detection ranged from 61% to 74%, below GM (87%), but contrasting with 47% for cultures. Combining PCR to EORTC/MSG criteria increased the sensitivity to 100%. Interestingly, tests performance in non-hematological patients ranged from 60% to 75%, and were higher than in hematological patients, and those with prior exposure to antifungals. All 16 isolates of Aspergillus fumigatus were susceptible; PCR did not detect any resistance marker in the 37 A. fumigatus PCR-positive samples. CONCLUSION: The molecular detection of Aspergillus directly in BAL samples greatly improved the diagnosis of IA, particularly in non-hematological patients.
OBJECTIVES: This study aimed to evaluate new tools to diagnose invasive aspergillosis (IA) directly from broncho-alveolar lavage (BAL) samples. METHODS: All consecutive patients with suspected IA who underwent bronchoscopy with BAL were prospectively included. Mycological culture and ELISA detection of galactomannan (GM) were performed on BAL. Two in-house and two marketed PCR assays were used on BAL DNA extracts to detect Aspergillus species. Susceptibility testing was performed after culture; marketed PCR assays detected mutations in the CYP51A gene associated to resistance. RESULTS: Within 3 years, 1555 BAL samples were processed, including 413 samples from 387 immunosuppressed patients. IA diagnosis was no-IA, possible, probable or proven IA in 326, 23, 37 and 1 patients, respectively. PCR assays sensitivity for Aspergillus detection ranged from 61% to 74%, below GM (87%), but contrasting with 47% for cultures. Combining PCR to EORTC/MSG criteria increased the sensitivity to 100%. Interestingly, tests performance in non-hematological patients ranged from 60% to 75%, and were higher than in hematological patients, and those with prior exposure to antifungals. All 16 isolates of Aspergillus fumigatus were susceptible; PCR did not detect any resistance marker in the 37 A. fumigatus PCR-positive samples. CONCLUSION: The molecular detection of Aspergillus directly in BAL samples greatly improved the diagnosis of IA, particularly in non-hematological patients.
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