title: Biosensor incorporating cell barrier architectures for detecting Staphylococcus aureus alpha toxin.
abstract: Alpha toxin is a common virulent factor of Staphylococcus aureus and is believed to play crucial roles in pathogenicity induced by S. aureus. Alpha toxin is also known to induce permeability to endothelial cell monolayers in vitro due to the formation of interendothelial gaps. The present study is directed towards measuring alpha toxin using a whole-cell-based biosensor. The biosensor, consisting of a confluent monolayer of human umbilical vein endothelial cells (HUVECs) on a potassium ion-selective electrode, takes advantage of cell permeability dysfunction to detect the presence of small quantities of alpha toxin. When a confluent monolayer of cells was formed on the membrane surface, the response of the electrode toward the marker ion, potassium, was inhibited. Upon exposing this sensor to varying concentrations of alpha toxin for 20 min, an increase in sensor response to potassium was observed. The response thus obtained was indirectly related to the concentration of alpha toxin. The detection limit of this sensor for alpha toxin was found to be 0.1 ng/ml. Cell monolayers were stained with silver nitrate to quantify the formation of intercellular gaps as well as to study the effect of this toxin on HUVECs morphology. A strong positive correlation was observed between the response obtained from the biosensor and the area of the intercellular gaps. Silver staining also revealed the tendency of cells to round up upon being exposed to alpha toxin.
authors.affiliation: Department of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40506, USA.
authors.forename: Gargi
authors.initials: G
authors.lastname: Ghosh
authors.forename: Leonidas G
authors.initials: LG
authors.lastname: Bachas
authors.forename: Kimberly W
authors.initials: KW
authors.lastname: Anderson
mesh_terms: D001427:Bacterial Toxins / Q000032:analysis* / Q000494:pharmacology; D015374:Biosensing Techniques / Q000379:methods*; D002463:Cell Membrane Permeability / Q000187:drug effects*; D042783:Endothelial Cells / Q000166:cytology; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D017736:Ion-Selective Electrodes; D011188:Potassium / Q000032:analysis; D013211:Staphylococcus aureus / Q000472:pathogenicity*; D010738:Type C Phospholipases / Q000032:analysis* / Q000494:pharmacology
chemical_list: D001427:Bacterial Toxins; D010738:Type C Phospholipases; D011188:Potassium