Electrical protein array chips for the detection of staphylococcal virulence factors.
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.
Quiel
Annett
A
Pharmazeutische Biotechnologie, Institut für Pharmazie, Ernst-Moritz-Arndt-Universität, 17487 Greifswald, Germany.
Jürgen
Britta
B
Piechotta
Gundula
G
Le Foll
Anne-Pascale
AP
Ziebandt
Anne-Kathrin
AK
Kohler
Christian
C
Köster
Daniela
D
Engelmann
Susanne
S
Erck
Christian
C
Hintsche
Rainer
R
Wehland
Jürgen
J
Hecker
Michael
M
Schweder
Thomas
T
D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis / Q000276:immunology; D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000032:analysis* / Q000276:immunology; D015374:Biosensing Techniques; D004563:Electrochemistry / Q000379:methods; D004768:Enterotoxins / Q000032:analysis* / Q000276:immunology / Q000652:urine; D005506:Food Contamination; D005516:Food Microbiology; D008892:Milk / Q000737:chemistry; D040081:Protein Array Analysis; D013211:Staphylococcus aureus / Q000737:chemistry*; D013212:Staphylococcus epidermidis / Q000276:immunology*; D037521:Virulence Factors / Q000032:analysis* / Q000276:immunology / Q000652:urine
D000907:Antibodies, Bacterial; D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D004768:Enterotoxins; C417673:IsaA antigen, staphylococcus; D037521:Virulence Factors; C031818:enterotoxin B, staphylococcal