Flower-Shaped PCR Scaffold-Based Lateral Flow Bioassay for Bacillus cereus Endospores Detection

Area of Science:

• Food safety
• Biosensor technology
• Pathogen detection

Background:

• Bacillus cereus endospores are difficult to detect using conventional methods.
• Existing detection techniques lack sensitivity and cost-effectiveness.
• There is a need for rapid, sensitive, and affordable pathogen detection systems.

Purpose of the Study:

• To develop a novel Flower-Shaped PCR Scaffold-based Lateral Flow Biosensor (FSPCRS-LFB) for detecting Bacillus cereus endospores.
• To evaluate the sensitivity, specificity, and cost-efficiency of the FSPCRS-LFB.
• To compare the FSPCRS-LFB with existing detection methods.

Main Methods:

• Development of an aptamer-integrated PCR scaffold as capture probes.
• Integration of the scaffold into a lateral flow biosensor platform.
• Validation of the FSPCRS-LFB using bacterial suspensions, endospore-cell mixtures, and real food samples (chicken and tea).

Main Results:

• The FSPCRS-LFB achieved a limit of detection (LOD) of 4.57 endospores/mL.
• Visual LOD was 10^2 endospores/mL, and LOD for endospore-cell mixtures was 6.78 CFU/mL.
• In food samples, LODs ranged from 52.8 to 74.5 endospores/mL with high recovery rates (82.19%–97.88%).
• The FSPCRS-LFB showed 3.7-fold higher sensitivity and was 26% cheaper than the streptavidin-biotin strategy and 87.5% cheaper than rolling circle amplification.

Conclusions:

• The FSPCRS-LFB is a highly sensitive and cost-effective biosensor for detecting Bacillus cereus endospores.
• This technology offers a rapid and reliable solution for point-of-care testing (POCT) in food safety.
• The FSPCRS-LFB presents a novel approach for pathogen detection, enhancing current capabilities.