Electrical protein array chips for the detection of staphylococcal virulence factors

• Published in: Applied microbiology and biotechnology 2010-02, Vol.85 (5), p.1619-1627
• Main Authors: Quiel, Annett, Jürgen, Britta, Piechotta, Gundula, Le Foll, Anne-Pascale, Ziebandt, Anne-Kathrin, Kohler, Christian, Köster, Daniela, Engelmann, Susanne, Erck, Christian, Hintsche, Rainer, Wehland, Jürgen, Hecker, Michael, Schweder, Thomas
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Bacterial - analysis; Antibodies, Bacterial - immunology; Antibodies, Immobilized; Antibodies, Monoclonal - immunology; Antigens; Antigens, Bacterial - analysis; Antigens, Bacterial - immunology; Biological and medical sciences; Biomedical and Life Sciences; Biosensing Techniques; Biotechnology; Collections; Electrochemistry - methods; Enterotoxins - analysis; Enterotoxins - immunology; Enterotoxins - urine; Enzymes; Food Contamination; Food contamination & poisoning; Food Microbiology; Fundamental and applied biological sciences. Psychology; Immunoassay; Life Sciences; Medical diagnosis; Methods; Microbial Genetics and Genomics; Microbiology; Milk - chemistry; Monoclonal antibodies; Nosocomial infections; Phosphatase; Protein Array Analysis; Proteins; R&D; Research & development; Semiconductors; Staphylococcus aureus; Staphylococcus aureus - chemistry; Staphylococcus epidermidis; Staphylococcus epidermidis - immunology; Staphylococcus infections; Studies; Virulence; Virulence Factors - analysis; Virulence Factors - immunology; Virulence Factors - urine; Viruses
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-009-2347-3

A new approach for the detection of virulence factors of Staphylococcus aureus and Staphylococcus epidermidis using an electrical protein array chip technology is presented. The procedure is based on an enzyme-linked sandwich immunoassay, which includes recognition and binding of virulence factors by specific capture and detection antibodies. Detection of antibody-bound virulence factors is achieved by measuring the electrical current generated by redox recycling of an enzymatically released substance. The current (measured in nanoampere) corresponds to the amount of the target molecule in the analyzed sample. The electrical protein chip allows for a fast detection of Staphylococcus enterotoxin B (SEB) of S. aureus and immunodominant antigen A homologue (IsaA homologue) of S. epidermidis in different liquid matrices. The S. aureus SEB virulence factor could be detected in minimal medium, milk, and urine in a concentration of 1 ng/ml within less than 23 min. Furthermore, a simultaneous detection of SEB of S. aureus and IsaA homologue of S. epidermidis in a single assay could be demonstrated.