Biofunctionalized Magnetic Nanoparticles with Multiplex Touchdown PCR for Simultaneous and Rapid Detection/Identification of Campylobacter jejuni and Campylobacter coli

The simple, accurate, and rapid detection of foodborne pathogens is essential for public health. Development an immunomagnetic separation (IMS) multiplex touchdown PCR (IMS–multiplex TD–PCR) assay simultaneous distinguishing C. jejuni coli reported herein. Polyclonal antibody (pAb) against multiepitope antigen (MEA) was conjugated to ferromagnetic nanoparticles (FMNs) produce anti-MEA FMNs. Optimal FMNs loading yielded 26.7 μg immunoglobulin G (IgG) molecules per mg with average size 72 ± 9 nm, corresponding 83% rate pAb conjugation. Anti-MEA (20 μg) IMS captured culturable cells at id="M2"> 3.54 × 10 2 colony-forming unit (CFU)/mL in pure culture, while higher amounts (40 60 reduced the recovery. scanning electron microscope (SEM) analysis revealed attachment target bacteria, forming aggregated magnetic ellipse-like shapes. subsequent TD–PCR simultaneously detected distinguished 104 CFU/mL mixed culture 103 each individual species. Furthermore, limit (LOD) IMS–multiplex CFU/g spiked chicken breast samples. Specificity 100% both as none amplicons were control samples where Campylobacter absent. This able detect distinguish a time result 4 h without enrichment step, making it promising approach culture-free products.