Gold nanoparticle-based enhanced chemiluminescence immunosensor for detection of Staphylococcal Enterotoxin B (SEB) in food.
Staphylococcal enterotoxins (SEs) are major cause of foodborne diseases, so sensitive detection (<1 ng/ml) methods are needed for SE detection in food. The surface area, geometric and physical properties of gold nanoparticles make them well-suited for enhancing interactions with biological molecules in assays. To take advantage of the properties of gold nanoparticles for immunodetection, we have developed a gold nanoparticle-based enhanced chemiluminescence (ECL) immunosensor for detection of Staphylococcal Enterotoxin B (SEB) in food. Anti-SEB primary antibodies were immobilized onto a gold nanoparticle surface through physical adsorption and then the antibody-gold nanoparticle mixture was immobilized onto a polycarbonate surface. SEB was detected by a "sandwich-type" ELISA assay on the polycarbonate surface with a secondary antibody and ECL detection. The signal from ECL was read using a point-of-care detector based on a cooled charge-coupled device (CCD) sensor or a plate reader. The system was used to test for SEB in buffer and various foods (mushrooms, tomatoes, and baby food meat). The limit of detection was found to be approximately 0.01 ng/mL, which is approximately 10 times more sensitive than traditional ELISA. The gold nanoparticles were relatively easy to use for antibody immobilization because of their physical adsorption mechanism; no other reagents were required for immobilization. The use of our simple and inexpensive detector combined with the gold nanoparticle-based ECL method described here is adaptable to simplify and increase sensitivity of any immunological assay and for point-of-care diagnostics.
Yang
Minghui
M
Center for Advanced Sensor Technology, University of Maryland Baltimore County, MD 21250, USA.
Kostov
Yordan
Y
Bruck
Hugh A
HA
Rasooly
Avraham
A
D000327:Adsorption; D000906:Antibodies; D001431:Bacteriological Techniques / Q000379:methods; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004768:Enterotoxins / Q000032:analysis* / Q000276:immunology; D005516:Food Microbiology; D006046:Gold / Q000276:immunology; D049449:Luminescence; D053768:Metal Nanoparticles; D011076:Polycarboxylate Cement
D000906:Antibodies; D004768:Enterotoxins; D011076:Polycarboxylate Cement; C005506:polycarbonate; C031818:enterotoxin B, staphylococcal; D006046:Gold