title: Immunoreactive proteins of Saccharopolyspora rectivirgula for farmer's lung serodiagnosis.
abstract: OBJECTIVE
  Saccharopolyspora rectivirgula is the principal cause of farmer's lung disease (FLD). Serodiagnosis is based on immunoprecipitation techniques or enzyme immunoassays with homemade crude antigens and is not standardized. We aimed to produce specific recombinant antigens for the development of a standardized ELISA.
  
  
  METHODS
  We recruited 41 patients and 43 healthy exposed controls from five university hospital pneumology departments in France and Switzerland. S. rectivirgula proteins were extracted, separated by 2D electrophoresis, and subjected to Western blotting, with sera from FLD patients or controls. FLD-specific proteins were identified by MS and were produced as recombinant antigens. The diagnostic performance of ELISA tests using the recombinant antigens was assessed with all the sera from FLD patients and controls.
  
  
  RESULTS
  We identified 25 FLD-specific proteins, some of which play important roles in transport, nutrition, or virulence. We produced 17 of these proteins as recombinant antigens and assessed their suitability for inclusion in the ELISA test. A combination of three of these proteins (SR1FA, SR17, and SR22) proved remarkably effective at discriminating between patients and controls, with a sensitivity of 83% and a specificity of 77%.
  
  
  CONCLUSIONS
  The recombinant antigens produced in this study constitute a major step toward the improvement of diagnostic performance and the standardization of FLD serodiagnosis.
authors.affiliation: UMR6249 Chrono-environnement, University of Franche-Comté, Besançon, France; Parasitology-Mycology Department, University Hospital of Besançon, Besançon, France.
authors.forename: Coralie
authors.initials: C
authors.lastname: Barrera
authors.forename: Laurence
authors.initials: L
authors.lastname: Millon
authors.forename: Bénédicte
authors.initials: B
authors.lastname: Rognon
authors.forename: Manfredo
authors.initials: M
authors.lastname: Quadroni
authors.forename: Sandrine
authors.initials: S
authors.lastname: Roussel
authors.forename: Jean-Charles
authors.initials: JC
authors.lastname: Dalphin
authors.forename: Isabelle
authors.initials: I
authors.lastname: Court-Fortune
authors.forename: Denis
authors.initials: D
authors.lastname: Caillaud
authors.forename: Stephane
authors.initials: S
authors.lastname: Jouneau
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Fellrath
authors.forename: Christophe
authors.initials: C
authors.lastname: Zaugg
authors.forename: Gabriel
authors.initials: G
authors.lastname: Reboux
authors.forename: Michel
authors.initials: M
authors.lastname: Monod
mesh_terms: D000328:Adult; D000368:Aged; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D005203:Farmer's Lung / Q000175:diagnosis / Q000276:immunology* / Q000382:microbiology; D005260:Female; D016908:Gram-Positive Bacterial Infections / Q000175:diagnosis / Q000276:immunology* / Q000382:microbiology; D054884:Host-Pathogen Interactions / Q000276:immunology; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D020543:Proteome / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D016930:Saccharopolyspora / Q000276:immunology* / Q000378:metabolism / Q000502:physiology; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods
chemical_list: D001426:Bacterial Proteins; D020543:Proteome
keywords: ELISA; Hypersensitivity pneumonitis; Immunoproteomics; Recombinant antigens; Serodiagnosis