Molecular detection of fungal community in sputa from cystic fibrosis patients

Ref ID: 19178

Author:

E. Campano*, V. Friaza, E. Quintana, J. Dapena, J.M. Varela, E. Calderon, C. De la Horra

Author address:

Seville, ES

Full conference title:

23rd European Congress of Clinical Microbiology and
Infectious Diseases

Date: 27 April 2014

Abstract:

Background: Cystic fibrosis is a hereditary disease characterized by defective mucociliary clearance and airway obstruction caused by a complex microbiota infection. Dus the polimicrobial nature of infections in patients with CF, the knowledge of airway microbiota is necessary. Several studies based on metagenomic tools have established the presence of bacterial pathogens in the lung of CF patients. Moreover, the presence of fungi is probably underestimated by the failure of culture methods. The emergence of fungal colonization is being studied by molecular approachs.
Objective. To describe the fungal community associated to CF patients by metagenomic tools
Methodology: The study enrolled 50 CF patients. DNA was isolated from spontaneous sputum using Nucleo Spin Kit. Fungal identification was performed by standard microbiological culture and molecular characterization by nested PCR to amplify ITS regions. The amplicons were sequenced and cloned in case of mix. The sequences were compared by BLAST search alignment on NCBI database to determinate phylogenetic affiliation. P. jirovecii identification was performed by real time PCR at mt LSU rRNA gene. Mixed infection with P. aeruginosa was detected by PCR.
Results: Fungal community has been identified in 32% of patients by standard methods: C. albicans, C parapsilosis and Aspergillus were the most represented fungus.
The molecular methods allow us to identify fungal species in 72% of CF patients. Among them, C.albicans was the most predominant isolated (45,4%). Secondly C.parapsilosis was isolated in 36,6% of cases. Interestingly, P. jirovecii was identified in 32% of patients. Several species of Aspergillus ssp were also isolated (17%).
Aditionally, a high rate of uncultured fungi and sequences corresponding to Unknown species with high phylogenetic specificity to fungi; other fungi such as several Cryptococcus sp, uncultured Lewia, C glabrata or Cladosporium sp were detected. Protozooa such as the genus Rhabostyla and Peritritia were also identified. These data of mixed colonization show a frequent association between P. aeruginosa/C. albicans and/or P. jirovecii; coinfection between fungi was also detected.
Conclusion: The use of new molecular tools designed to identify the microbiome of CF patients allow us to describe the whole airway fungi community. This approach is useful to detect emerging pathogens and a possible interaction between microorganisms in the context of Cystic fibrosis disorder

Abstract Number: P1081

Conference Year: 2013

Link to conference website: http://registration.akm.ch/einsicht.php?XNABSTRACT_ID=166087&XNSPRACHE_ID=2&XNKONGRESS_ID=180&XNMASK

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