title: Direct, Specific and Rapid Detection of Staphylococcal Proteins and Exotoxins Using a Multiplex Antibody Microarray.
abstract: BACKGROUND
  S. aureus is a pathogen in humans and animals that harbors a wide variety of virulence factors and resistance genes. This bacterium can cause a wide range of mild to life-threatening diseases. In the latter case, fast diagnostic procedures are important. In routine diagnostic laboratories, several genotypic and phenotypic methods are available to identify S. aureus strains and determine their resistances. However, there is a demand for multiplex routine diagnostic tests to directly detect staphylococcal toxins and proteins.
  
  
  METHODS
  In this study, an antibody microarray based assay was established and validated for the rapid detection of staphylococcal markers and exotoxins. The following targets were included: staphylococcal protein A, penicillin binding protein 2a, alpha- and beta-hemolysins, Panton Valentine leukocidin, toxic shock syndrome toxin, enterotoxins A and B as well as staphylokinase. All were detected simultaneously within a single experiment, starting from a clonal culture on standard media. The detection of bound proteins was performed using a new fluorescence reading device for microarrays.
  
  
  RESULTS
  110 reference strains and clinical isolates were analyzed using this assay, with a DNA microarray for genotypic characterization performed in parallel. The results showed a general high concordance of genotypic and phenotypic data. However, genotypic analysis found the hla gene present in all S. aureus isolates but its expression under given conditions depended on the clonal complex affiliation of the actual isolate.
  
  
  CONCLUSIONS
  The multiplex antibody assay described herein allowed a rapid and reliable detection of clinically relevant staphylococcal toxins as well as resistance- and species-specific markers.
authors.affiliation: Alere Technologies GmbH, Jena, Germany.|Institute for Medical Microbiology and Hygiene, Technische Universität Dresden, Dresden, Germany.|Infectognostics Forschungscampus Jena, Jena, Germany.
authors.forename: Bettina
authors.initials: B
authors.lastname: Stieber
authors.affiliation: Alere Technologies GmbH, Jena, Germany.|Institute for Medical Microbiology and Hygiene, Technische Universität Dresden, Dresden, Germany.|Infectognostics Forschungscampus Jena, Jena, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Monecke
authors.affiliation: Alere Technologies GmbH, Jena, Germany.|Infectognostics Forschungscampus Jena, Jena, Germany.
authors.forename: Elke
authors.initials: E
authors.lastname: Müller
authors.affiliation: Alere San Diego, Inc., San Diego, California, United States of America.
authors.forename: Joseph
authors.initials: J
authors.lastname: Büchler
authors.affiliation: Alere Technologies GmbH, Jena, Germany.|Infectognostics Forschungscampus Jena, Jena, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Ehricht
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D005098:Exotoxins / Q000276:immunology / Q000378:metabolism*; D060005:Genotyping Techniques; D057230:Limit of Detection; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D013211:Staphylococcus aureus / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D015415:Biomarkers; D005098:Exotoxins