期刊
MYCOPATHOLOGIA
卷 183, 期 1, 页码 171-183出版社
SPRINGER
DOI: 10.1007/s11046-017-0185-x
关键词
Microbiome; Mycobiome; Cystic fibrosis; 16S rDNA; ITS2; UDS
类别
资金
- Assistance Publique-Hopitaux de Paris (public sector, bourse de mobilite)
- Pfizer France Pharmaceutical Division
- French Ministry of Health and Research (PHRC) [2006/1902]
- Centre Hospitalier Regional Universitaire de Lille
- association Vaincre la Mucoviscidose (MucoFong and Mucofong-ATF) [N8 2006/351]
- Pfizer France Pharmaceutical Division [2006/158]
Given the complexity of the airway microbiota in the respiratory tract of cystic fibrosis (CF) patients, it seems crucial to compile the most exhaustive and exact list of the microbial communities inhabiting CF airways. The aim of the present study was to compare the bacterial and fungal diversity of sputa from adult CF patients during non-exacerbation period by culture-based and molecular methods, and ultra-deep-sequencing (UDS). Sputum samples from four CF patients were cultured and analysed by DNA extractions followed by terminal restriction fragment length polymorphism analysis through resolution of bacterial ribosomal gene (rDNA) fragments, and cloning plus sequencing of part of fungal rRNA genes. These approaches were compared with UDS method targeting 16S rDNA gene and the internal transcribed spacer (ITS) 2 region of rDNA. A total of 27 bacterial and 18 fungal genera were detected from the four patients. Five (18%) and 3 (16%) genera were detected by culture for bacteria and fungi, respectively, 9 (33%) and 3 (16%) by first generation sequencing (FGS) methods, and 26 (96%) and 18 (100%) by UDS. The mean number of genera detected by UDS per patient was statistically higher than by culture or FGS methods. Patients with severe airway disease as assessed by standard spirometry exhibited a reduced fungal and bacterial diversity. UDS approach evaluates more extensively the diversity of fungal and bacterial flora compared with cultures. However, it currently remains difficult to routinely use UDS mainly because of the lack of standardization, and the current cost of this method.
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