3D Printed Chips Empower CRISPR for Rapid On-Site Detection of Foodborne Bacteria

Recently, Professor Ding Xiong’s team from Southeast University published significant research findings in the journal Advanced Science, successfully developing a deployable platform for the multiplex detection of foodborne bacteria. This platform ingeniously combines an enhanced CRISPR detection system with 3D printed microfluidic chips, providing a revolutionary solution for rapid food safety testing.

The team’s enhanced single-tube helicase-assisted RPA-double CRISPR/uAsCas12a (EOD-CRISPR) technology significantly improves detection speed, sensitivity, and fluorescence signal intensity by adding bovine serum albumin (BSA), helicase gp41, and uAsCas12a nuclease to the reaction system. This “one-pot method” design simplifies the operational process, laying the foundation for on-site applications.

Meanwhile, the research employs 3D printing technology to create a microfluidic disk, integrating four central chambers and eight outer chambers, supporting the simultaneous detection of four target bacteria. By embedding glass fiber membranes in the chambers and adding 5% trehalose and 1% glycine as protectants, the stable lyophilization of EOD-CRISPR reagents was successfully achieved, addressing the challenges of reagent storage and transportation.

Ultimately, the system integrates a 3D printed DNA extractor, a portable fluorescence detector, and a mini centrifuge with lyophilized chips, constructing a complete on-site detection platform. Sample processing to result reading can be completed in a short time without the need for complex laboratory equipment.

Performance Validation and Application Value

Experimental results indicate that the platform’s detection sensitivity for Bacillus cereus, Salmonella enterica, Staphylococcus aureus, and Escherichia coli O157:H7 is comparable to standard PCR methods, with excellent specificity and no cross-reactivity. It demonstrated stable performance in simulated contaminated food sample testing, confirming its immense potential for on-site food safety detection.

Future Prospects

This technology not only provides an efficient tool for detecting foodborne bacteria but can also be extended to clinical diagnostics and animal disease prevention in the future. The research team stated that they will further optimize DNA extraction efficiency to promote the platform’s practical application in more scenarios, safeguarding public health.

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