A CRISPR-Cas12a-powered magnetic relaxation switching biosensor for the sensitive detection of Salmonella

Magnetic relaxation switching (MRS) biosensors are attractive in the field of food safety owing to their simplicity and high signal-to-noise ratio. But they are less in sensitivity and stability caused by the insufficient crosslinking or non-specific binding of magnetic nanoparticles (MNPs) with targets. To address this problem, the CRISPR-Cas12a system was introduced into an MRS biosensor for the first time, to precisely control the binding of two types of MNPs with sizes of 130 nm (MNP₁₃₀) and 30 nm (MNP₃₀), for the sensitive detection of Salmonella. Delicately, the biosensor was designed based on the different magnetic properties of the two sizes of MNPs. The target Salmonella activated the collateral cleavage activity of the CRISPR-Cas12a system, which inhibited the binding of the two sizes of MNPs, resulting in an increase of unbound MNP₃₀. After separating MNP₁₃₀-MNP₃₀ complexes and MNP₁₃₀ from MNP₃₀, the free MNP₃₀ left in solution acted as transverse relaxation time (T₂) signal reporters for Salmonella detection. Under optimized conditions, the CRISPR-MRS biosensor presented a limit of detection of 1.3 × 10² CFU mL^-1 for Salmonella, which is lower than most MRS biosensor analogues. It also showed satisfactory specificity and performed well in spiked chicken meat samples. This biosensing strategy not only extends the reach of the CRISPR-Cas12a system in biosensors but also offers an alternative for pathogen detection with satisfactory sensitivity.