Toward the Standardization of Mycological Examination of Sputum Samples in Cystic Fibrosis : Results from a French Multicenter Prospective Study
Noémie Coron
(1, 2, 3)
,
Marc Pihet
(1)
,
Emilie Fréalle
(4)
,
Yolande Lemeille
(5, 6)
,
Claudine Pinel
(5, 6)
,
Hervé Pelloux
(5, 6)
,
Gilles Gargala
(7)
,
Loïc Favennec
(7)
,
Isabelle Accoceberry
(3, 8)
,
Isabelle Durand-Joly
(9)
,
Frédéric Dalle
(10)
,
Frédéric Huet
(10, 11)
,
Annlyse Fanton
(11, 10)
,
Amale Boldron
(9)
,
Guy-André Loeuille
(11, 9)
,
Philippe Domblides
(12, 3)
,
Bérengère Coltey
(13, 5)
,
Isabelle Pin
(5)
,
Catherine Llerena
(5)
,
Françoise Troussier
(1)
,
Christine Person
(1)
,
Christophe Marguet
(14, 15)
,
Nathalie Wizla
(16)
,
Caroline Thumerelle
(16)
,
Dominique Turck
(16)
,
Stéphanie Bui
(12, 17, 3)
,
Michael Fayon
(12, 17, 3)
,
Alain Duhamel
(18)
,
Anne Prévotat
(16)
,
Benoit Wallaert
(16)
,
Sylvie Leroy
(16, 19, 20)
,
Jean-Philippe Bouchara
(1)
,
Laurence Delhaes
(4, 2)
1
CHU Angers -
Centre Hospitalier Universitaire d'Angers
2 CRCTB - Centre de recherche Cardio-Thoracique de Bordeaux [Bordeaux]
3 CHU Bordeaux
4 Service de Parasitologie-Mycologie [CHRU LIlle]
5 CHUGA - Centre Hospitalier Universitaire [CHU Grenoble]
6 UGA [2016-2019] - Université Grenoble Alpes [2016-2019]
7 Hôpital Charles Nicolle [Rouen]
8 UB - Université de Bordeaux
9 CH Dunkerque
10 CHU Dijon - Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand
11 CRCM - Centre de Ressources et de Compétences de la Mucoviscidose [CHU Dijon]
12 CRCM - Centre de Recherche en Chimie Moléculaire
13 CRCM Adulte [Grenoble]
14 Service de pédiatrie médicale et médecine de l'adolescent [Rouen]
15 GRAM 1.0 - Groupe de Recherche sur les Antimicrobiens et les Micro-Organismes
16 CHRU Lille - Centre Hospitalier Régional Universitaire [CHU Lille]
17 CIC Bordeaux
18 CERIM - Centre d'Etudes et de Recherche en Informatique Médicale [Lille]
19 CHU - Hôpital Pasteur [Nice]
20 CHU Nice - Centre Hospitalier Universitaire de Nice
2 CRCTB - Centre de recherche Cardio-Thoracique de Bordeaux [Bordeaux]
3 CHU Bordeaux
4 Service de Parasitologie-Mycologie [CHRU LIlle]
5 CHUGA - Centre Hospitalier Universitaire [CHU Grenoble]
6 UGA [2016-2019] - Université Grenoble Alpes [2016-2019]
7 Hôpital Charles Nicolle [Rouen]
8 UB - Université de Bordeaux
9 CH Dunkerque
10 CHU Dijon - Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand
11 CRCM - Centre de Ressources et de Compétences de la Mucoviscidose [CHU Dijon]
12 CRCM - Centre de Recherche en Chimie Moléculaire
13 CRCM Adulte [Grenoble]
14 Service de pédiatrie médicale et médecine de l'adolescent [Rouen]
15 GRAM 1.0 - Groupe de Recherche sur les Antimicrobiens et les Micro-Organismes
16 CHRU Lille - Centre Hospitalier Régional Universitaire [CHU Lille]
17 CIC Bordeaux
18 CERIM - Centre d'Etudes et de Recherche en Informatique Médicale [Lille]
19 CHU - Hôpital Pasteur [Nice]
20 CHU Nice - Centre Hospitalier Universitaire de Nice
Emilie Fréalle
- Fonction : Auteur
- PersonId : 776972
- ORCID : 0000-0003-3129-979X
- IdRef : 069218005
Isabelle Pin
- Fonction : Auteur
- PersonId : 756749
- ORCID : 0000-0002-7529-6999
Christophe Marguet
- Fonction : Auteur
- PersonId : 930585
Caroline Thumerelle
- Fonction : Auteur
- PersonId : 984232
Dominique Turck
- Fonction : Auteur
- PersonId : 910333
Jean-Philippe Bouchara
- Fonction : Auteur
- PersonId : 920220
Laurence Delhaes
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- Fonction : Auteur correspondant
- PersonId : 955454
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Résumé
Fungal respiratory colonization of cystic fibrosis (CF) patients emerges as a new concern; however, the heterogeneity of mycological protocols limits investigations. We first aimed at setting up an efficient standardized protocol for mycological analysis of CF sputa that was assessed during a prospective, multicenter study: “MucoFong” program (PHRC-06/1902). Sputa from 243 CF patients from seven centers in France were collected over a 15-month period and submitted to a standardized protocol based on 6 semi-selective media. After mucolytic pretreatment, sputa were plated in parallel on cycloheximide-enriched (ACT37), erythritol-enriched (ERY37), benomyl dichloran–rose bengal (BENO37) and chromogenic (CAN37) media incubated at 37 °C and on Sabouraud–chloramphenicol (SAB27) and erythritol-enriched (ERY27) media incubated at 20–27 °C. Each plate was checked twice a week during 3 weeks. Fungi were conventionally identified; time for detection of fungal growth was noted for each species. Fungal prevalences and media performances were assessed; an optimal combination of media was determined using the Chi-squared automatic interaction detector method. At least one fungal species was isolated from 81% of sputa. Candida albicans was the most prevalent species (58.8%), followed by Aspergillus fumigatus (35.4%). Cultivation on CAN37, SAB27, ACT37 and ERY27 during 16 days provided an optimal combination, detecting C. albicans, A. fumigatus, Scedosporium apiospermum complex and Exophiala spp. with sensitivities of 96.5, 98.8, 100 and 100%. Combination of these four culture media is recommended to ensure the growth of key fungal pathogens in CF respiratory specimens. The use of such consensual protocol is of major interest for merging results from future epidemiological studies.