title: Characterization of the nasopharyngeal microbiota in health and during rhinovirus challenge.
abstract: BACKGROUND
  The bacterial communities of the nasopharynx play an important role in upper respiratory tract infections (URTIs). Our study represents the first survey of the nasopharynx during a known, controlled viral challenge. We aimed to gain a better understanding of the composition and dynamics of the nasopharyngeal microbiome during viral infection.
  
  
  METHODS
  Rhinovirus illnesses were induced by self-inoculation using the finger to nose or eye natural transmission route in ten otherwise healthy young adults. Nasal lavage fluid samples (NLF) samples were collected at specific time points before, during, and following experimental rhinovirus inoculation. Bacterial DNA from each sample (N = 97 from 10 subjects) was subjected to 16S rRNA sequencing by amplifying the V1-V2 hypervariable region followed by sequencing using the 454-FLX platform.
  
  
  RESULTS
  This survey of the nasopharyngeal microbiota revealed a highly complex microbial ecosystem. Taxonomic composition varied widely between subjects and between time points of the same subject. We also observed significantly higher diversity in not infected individuals compared to infected individuals. Two genera - Neisseria and Propionibacterium - differed significantly between infected and not infected individuals. Certain phyla, including Firmicutes, Actinobacteria, and Proteobacteria, were detected in all samples.
  
  
  CONCLUSIONS
  Our results reveal the complex and diverse nature of the nasopharyngeal microbiota in both healthy and viral-challenged adults. Although some phyla were common to all samples, differences in levels of diversity and selected phyla were detected between infected and uninfected participants. Deeper, species-level metagenomic sequencing in a larger sample is warranted.
authors.affiliation: Center for Public Health Genomics, University of Virginia, PO Box 800717, Charlottesville, USA ; Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, USA.
authors.forename: E Kaitlynn
authors.initials: EK
authors.lastname: Allen
authors.affiliation: Bioinformatics Core Facility, University of Virginia, Charlottesville, USA.
authors.forename: Alex F
authors.initials: AF
authors.lastname: Koeppel
authors.affiliation: Department of Pediatrics, University of Virginia, Charlottesville, USA.
authors.forename: J Owen
authors.initials: JO
authors.lastname: Hendley
authors.affiliation: Bioinformatics Core Facility, University of Virginia, Charlottesville, USA.
authors.forename: Stephen D
authors.initials: SD
authors.lastname: Turner
authors.affiliation: Department of Otolaryngology, University of Virginia, Charlottesville, USA.
authors.forename: Birgit
authors.initials: B
authors.lastname: Winther
authors.affiliation: Center for Public Health Genomics, University of Virginia, PO Box 800717, Charlottesville, USA ; Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, USA ; Department of Medicine, University of Virginia, Charlottesville, USA.
authors.forename: Michèle M
authors.initials: MM
authors.lastname: Sale
keywords: Microbiota; longitudinal; nasopharynx; rhinovirus illness; sequencing