title: Improving patient care via development of a protein-based diagnostic test for microbe-specific detection of chronic rhinosinusitis.
abstract: OBJECTIVE
  The hypothesis is that signature bacterial proteins can be identified in sinus secretions via high-throughput, proteomic based techniques. Nontypeable Haemophilus influenzae (NTHI) is the most common bacterial pathogen associated with sinusitis and serves as proof of principle pathogen for identifying biomarkers.
  
  
  METHODS
  In vitro and in vivo studies using proteomic-based analysis of cultures of NTHI and a novel, experimental chinchilla polymicrobial sinusitis model.
  
  
  METHODS
  Nano-liquid chromatography /tandem mass spectrometry (nano-LC-MS/MS) was performed to annotate the secretome from an NTHI biofilm. A model of NTHI-induced sinusitis was developed in a chinchilla, and NTHI proteins were detected in chinchilla secretions. A reference standard RT-PCR-based assay was adapted to allow for sensitivity and specificity testing of the identified signature biomarkers in human patients.
  
  
  RESULTS
  Outer membrane proteins P2 (OMP-P2) and P5 (OMP-P5) were identified as promising candidates for the detection of NTHI biofilms and positively detected in nasopharyngeal secretions of chinchillas experimentally infected with NTHI. An RT-PCR based test for the presence of NTHI biofilms demonstrated 100% sensitivity and 100% specificity when tested against eight unique strains commonly found in human bacterial rhinosinusitis.
  
  
  CONCLUSIONS
  Proteomic analysis was successful in identifying signature proteins for possible use as a biomarker for chronic rhinosinusitis (CRS). OMP-P2 and OMP-P5 were validated as promising candidates and were positively detected from nasopharyngeal secretions from chinchillas experimentally infected with NTHI. Collectively, these data support the use of OMP-P2 and OMP-P5 as biomarkers for a human clinical trial to develop a point-of-care medical diagnostic test to assist in the diagnosis and treatment of CRS.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, The Ohio State University, Columbus, Ohio, U.S.A; Center for Microbial Pathogenesis, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, U.S.A.
authors.forename: Subinoy
authors.initials: S
authors.lastname: Das
authors.forename: Lucia E
authors.initials: LE
authors.lastname: Rosas
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Jurcisek
authors.forename: Laura A
authors.initials: LA
authors.lastname: Novotny
authors.forename: Kari B
authors.initials: KB
authors.lastname: Green
authors.forename: Lauren O
authors.initials: LO
authors.lastname: Bakaletz
mesh_terms: D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D018441:Biofilms / Q000254:growth & development; D015415:Biomarkers / Q000378:metabolism; D002682:Chinchilla; D002908:Chronic Disease; D004195:Disease Models, Animal; D006192:Haemophilus Infections / Q000175:diagnosis* / Q000235:genetics; D006193:Haemophilus influenzae / Q000145:classification* / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D005791:Patient Care; D040901:Proteomics; D058996:Quality Improvement; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012015:Reference Standards; D015203:Reproducibility of Results; D012220:Rhinitis / Q000175:diagnosis* / Q000382:microbiology; D012680:Sensitivity and Specificity; D012852:Sinusitis / Q000175:diagnosis* / Q000382:microbiology; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D015415:Biomarkers; C074703:outer membrane protein P2, Haemophilus influenzae type b; C078167:OMPA outer membrane proteins
keywords: Sinusitis; bacterial; diagnostic; point-of-care; proteomic
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