Cell Based-Green Fluorescent Biosensor Using Cytotoxic Pathway for Bacterial Lipopolysaccharide Recognition.

Area of Science:

• Biotechnology
• Biosensor Development
• Molecular Biology

Background:

• Lipopolysaccharide (LPS) is a key biomarker for Gram-negative bacterial contamination.
• Accurate and efficient detection of LPS is crucial for public health and food safety.

Purpose of the Study:

• To develop a novel, user-friendly, and highly efficient cell-based fluorescent biosensor for LPS detection.
• To utilize a recombinant plasmid in engineered cells to enable optical detection of LPS.

Main Methods:

• Constructed a recombinant plasmid combining the ZC3H12A promoter and enhanced green fluorescence protein (EGFP).
• Transfected 293/hTLR4A-MD2-CD14 cells with the plasmid for LPS-induced fluorescence.
• Utilized a high-content screening imaging system for observation and analysis.

Main Results:

• The stable transfected cells exhibited green fluorescence upon LPS treatment, enabling direct observation.
• The biosensor demonstrated a low detection limit of 0.075 μg/mL for LPS.
• Successfully differentiated between Gram-negative and Gram-positive bacteria and detected LPS in fruit juices.

Conclusions:

• The developed cell-based fluorescent biosensor is effective for sensitive and high-throughput LPS detection.
• This biosensor shows significant potential for optical sensing in foodstuffs and biological products.
• Contributes to controlling foodborne diseases and ensuring public food safety through rapid bacterial identification.