Sensitive, Rapid, Quantitative and in Vitro Method for the Detection of Biologically Active Staphylococcal...

Area of Science:

• Microbiology and Immunology
• Food Safety and Toxicology

Background:

• Staphylococcus aureus produces enterotoxins (SEs) causing foodborne illnesses and T cell activation.
• Staphylococcal enterotoxin E (SEE) is implicated in food poisoning outbreaks globally.
• Current detection methods (animal bioassays) are insensitive, costly, and ethically concerning.

Purpose of the Study:

• To develop rapid, sensitive, and quantitative bioassays for active Staphylococcal enterotoxin E (SEE).
• To establish a reliable alternative to animal-based detection methods.

Main Methods:

• Utilized a genetically engineered T cell-line with a luciferase reporter gene.
• Combined engineered T cells with Raji B cells presenting SEE-MHC class II complexes.
• Measured bioluminescence in a dose-dependent manner upon SEE exposure.

Main Results:

• Achieved a limit of detection for biologically active SEE at 1 fg/mL.
• Demonstrated a 6-log dynamic range for bioluminescence readout.
• The new bioassay is 10⁸ times more sensitive than traditional animal bioassays.

Conclusions:

• The developed T cell-based bioassay is highly sensitive and quantitative for SEE detection.
• This method provides a superior alternative to animal bioassays for food safety applications.
• Enables rapid and ethical monitoring of Staphylococcal enterotoxin E contamination.