Aspergillus Azole resistance. Mecanisms of resistance. Origin of resistance
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1 Logo de la journée ici Etude prospective multicentrique (RADAR) de la résistance d Aspergillus fumigatus aux azolés en France J.B. MURAT 1,2, F. CHOUKRI 1, J.M. COSTA 3, I. ACCOCEBERRY 4, C. ANGEBAULT 5, M.E. BOUGNOUX 5, N. BOURGEOIS 6, M. CORNET 7, L. DELHAES 8, N. FAUCHET 9, O. FAURE 7, A. FEKKAR 10, J. GUITARD 11, C. HENNEQUIN 11, L. LACHAUD 12, A. PAUGAM 13, R. KIM 2, E. SITTERLE 1, J. GUILLOT 1, F. BOTTEREL 1,2, E. DANNAOUI 1,14. 1 Dynamyc, UPEC, ENVA, 2 Hosp. Mondor, Créteil, 3 Lab. CERBA, 4 Hosp. Pellegrin, Bordeaux, 5 Hosp. Necker-Enfants malades, Paris, 6 CHRU, Montpellier, 7 CHU, Grenoble, 8 CHU, Lille, 9 CHIC, Créteil, 10 Hosp. Pitié, Paris, 11 Hosp. Saint-Antoine, Paris, 12 Hosp. Caremeau, Nimes, 13 Hosp. Cochin, Paris, 14 Hosp. Pompidou, Paris, France.
2 Aspergillus and aspergillosis Filamentous fungus Soil-borne microorganism A. fumigatus: the most common +++ Aspergillus heads numerous spores Infection - Treatment spores inhalation pulmonary disease Aspergillose pulmonaire invasive (API) Aspergillome (balle fongique) Aspergillose brocho-pulmonaire allergique (ABPA) Voriconazole +++ Dagenais, T. R., Clin Microbiol Rev 22:
3 Aspergillus Azole resistance Emergence of resistance Worldwide distribution Detection of resistance in clinical samples (irrespective to mecanism) USA The Netherlands Germany Norway Spain Denmark UK France Sweden Belgium Greece Italy Austria Iran India China Japan Snelders et al PLoS Med. 5:e219 Australia
4 Aspergillus Azole resistance Mecanisms of resistance Origin of resistance
5 Azole resistance - France Aspergillus Azole resistance Detection of resistance in clinical samples (irrespective to mecanism) Few data First detection in the late 1990s 1,2 Few data 3-6 Mostly in selected populations Retrospective analysis 7 1 year (2012) all patients o Prevalence 1.8% (isolates) 1. Dannaoui, E., et al JAC 44, Dannaoui, E., et al JAC 47, Morio, F., JAC 67: Burgel, P. R., AAC 56: Rocchi, S. 2014, JCM 52, Lavergne RA., 2015, AAC Apr Choukri, F., et al Med Mycol, in Press 8. Choukri, F., et al TIMM Retrospective analysis 8 12 years ( ) Lung Tx patients o Prevalence 8.9% (isolates)
6 Aim Aim of the study The RADAR prospective study Risk in Aspergillosis: Detection of Azole Resistance Surveillance study Evaluation of prevalence and mechanisms C Prospective - Multicenter All Aspergillus spp. screened on azole-containing plates ITZ VRZ
7 RADAR Lille Laurence Delhaes Bordeaux Isabelle Accoceberry Nîmes Laurence Lachaud Montpellier Nathalie Bourgeois Grenoble Odile Faure Muriel Cornet Necker Cecile Angebault Marie-Elisabeth Bougnoux Pitié Arnaud Fekkar Cochin André Paugam Saint-Antoine Juliette Guitard Christophe Hennequin HEGP Eric Dannaoui HMN Françoise Botterel CHIC Nathalie Fauchet
8 RADAR study Study design Screening 1 year Standardizedprotocol Meeting pre-study Video QC exercise Video All samples 1 colony Incubation 48h 37 C
9 Methods Methods Screening 12 centers Resistant isolates EUCAST Isolates centralized in one lab Resistant isolates Species identification (Beta-tubulin) Mutations (Cyp51 sequencing)
10 Screening results Number of isolates Total isolates screened expected screened real A. fumigatus resistant isolates Resistant Af expected Resistant Af real 0 août-13 nov.-13 mars-14 juin-14 sept.-14 déc.-14
11 Screening results Number by center Distribution of species Number of isolates N= Center # A. fumigatus A. niger A. flavus A. nidulans A. terreus A. versicolor Other
12 The RADAR prospective study Screening results A. fumigatus R Resistant Screened
13 PRELIMINARY results A. fumigatus resitant isolates - Analysis in progress EUCAST N= R/I 24 S 2 N. pseudofisheri 108 A. fumigatus Prevalence: 4.2%
14 Results TR 34 /L98H mutation 108 A. fumigatus resistant isolates The RADAR prospective study Cyp51 mutation n Cyp51 mutation n TR 34 /L98H 66 G448S 1 Pas de mutation 19 G54W 1 G54R 5 M220V 1 TR 34 /L98H/H147Y 5 TR 34 /L98H/S297T/F495I 1 H285Y 3 TR 46 /Y121F/T289A 1 M220I 2 Y121F 1 Various underlying diseases CF SOT Kc Hematol malignancies Various aspergillosis Colonization ABPA IA Aome
15 Conclusions Our results provide new data about azole resistance in France Azole resistant A. fumigatus isolates can be recovered in France from patients with different underlying diseases The TR 34 /L98H mutation is the most frequent Cyp51A mutation involved AFST of Aspergillus spp. is recommended and further surveillance epidemiological studies are needed
16 Thank you
17 Epidemiology Unresolved issues Problem of resistance detection Available techniques EUCAST Culture positive samples Screening agar plates / Etest Cyp51 mutation Culture negative samples PCR (Mycogenie, Aspergenius) Not all mutations Pb of sensitivity NGS
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Received 5 June 2009/Returned for modification 31 July 2009/Accepted 23 October 2009
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