title: Microbiome complexity and Staphylococcus aureus in chronic rhinosinusitis.
abstract: OBJECTIVE
  The aim of this study was to compare microbiological culture-based and culture-independent (16S rRNA gene sequencing) methodologies for pathogen identification in chronic rhinosinusitis (CRS) patients. We hypothesized that bacterial culture and DNA sequencing would yield largely concurrent results, although sequencing would detect greater bacterial diversity, and the sinonasal microbiomes of CRS patients would differ in composition and diversity compared with non-CRS controls.
  
  
  METHODS
  Cross-sectional observational study.
  
  
  METHODS
  Middle meatus swabs from CRS patients collected during endoscopic sinus surgery were analyzed by both clinical culture and broad-range analysis of 16S rRNA gene pyrosequences.
  
  
  RESULTS
  A total of 21 swab samples from 15 CRS patients and five non-CRS controls were analyzed. One CRS patient was also swabbed 3 weeks postoperatively due to evidence of purulence during a clinical visit. All subjects had positive bacterial cultures, with a mean of 2.8 isolates per subject. The most prevalent cultivars were coagulase-negative staphylococci (15/20 specimens, 75%), Staphylococcus aureus (10/20, 50%), and Propionibacterium acnes (6/20, 30%). Among 57,407 pyrosequences generated, the most prevalent were from coagulase-negative staphylococci (21/21 specimens, 100%), Corynebacterium spp (18/21, 85.7%), P acnes (16/21, 76.2%), and S aureus (14/21, 66.7%). Bacterial diversity correlated with recent antibiotic use, asthma, prior sinus surgery, and relative abundance of S aureus.
  
  
  CONCLUSIONS
  DNA pyrosequencing revealed greater biodiversity than culture, although in most cases culture results represented a subset of the abundant DNA sequence types. CRS patients were characterized by altered microbial composition (P = .02) and greater abundance of S aureus (P = .03).
authors.affiliation: Division of Infectious Diseases, University of Colorado School of Medicine, Aurora, Colorado, USA.
authors.forename: Leah M
authors.initials: LM
authors.lastname: Feazel
authors.forename: Charles E
authors.initials: CE
authors.lastname: Robertson
authors.forename: Vijay R
authors.initials: VR
authors.lastname: Ramakrishnan
authors.forename: Daniel N
authors.initials: DN
authors.lastname: Frank
mesh_terms: D002908:Chronic Disease; D003430:Cross-Sectional Studies; D005260:Female; D006801:Humans; D008297:Male; D054892:Metagenome; D008875:Middle Aged; D012329:RNA, Bacterial / Q000235:genetics*; D012189:Retrospective Studies; D012220:Rhinitis / Q000150:complications / Q000382:microbiology*; D017423:Sequence Analysis, RNA; D012852:Sinusitis / Q000150:complications / Q000382:microbiology*; D013203:Staphylococcal Infections / Q000382:microbiology*; D013211:Staphylococcus aureus / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D012329:RNA, Bacterial