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In this study, we described a novel impedance biosensor combining immunomagnetic separation with urease catalysis for sensitive detection of foodborne bacteria using Listeria monocytogenes as model and an immobilization-free microelectrode as detector. The monoclonal antibodies (MAbs) were immobilized on the surface of the magnetic nanoparticles (MNPs) with the diameter of 180 nm by biotin–streptavidin system for specifically and efficiently separating Listeria cells from sample background. The polyclonal antibodies (PAbs) and the urease were modified onto the surface of the gold nanoparticles (AuNPs) with the diameter of 20 nm and the modified AuNPs were used to react with Listera to form the MNP–MAb– Listeria –PAb–AuNP–urease sandwich complexes. The urease in the complexes could catalyze the hydrolysis of the urea into ammonium carbonate and this led to an increase in the ionic strength of the media, which could be detected by the microelectrode. The magnetic separation efficiencies for L. monocytogenes at the concentrations ranging from 3.0×10 1 to 3.0×10 4 CFU/mL were over 95% for the pure cultures and over 85% for the spiked lettuce samples. The lower detection limit of this biosensor for L. monocytogenes was found to be 300 CFU/mL in both the pure cultures and the spiked lettuce samples. The microelectrode was demonstrated to be reusable for over 50 times with thorough cleaning by deionized water. This biosensor showed its potential to provide a simple, low-cost and sensitive method for rapid screening of foodborne pathogens and could be extended for detection of other biological or chemical targets. [ABSTRACT FROM AUTHOR]

Salmonella is the leading risk factor in food safety. Rapid, sensitive and accurate detection of Salmonella is a key to prevent and control the outbreaks of foodborne diseases caused by Salmonella. In this study, we reported a colorimetric biosensor for ultrasensitive detection of Salmonella Typhimurium using a magnetic grid separation column to efficiently separate target bacteria from large volume of sample and platinum loaded zeolitic imidazolate framework-8 (Pt@ZIF-8) nanocatalysts to effectively amplify biological signal. The target Salmonella cells in large volume of sample were first separated and concentrated using the magnetic grid separation column with immune magnetic particle chains, then conjugated with the immune Pt@ZIF-8 nanocatalysts to mimic peroxidase for catalysis of hydrogen peroxide-3,3′,5,5′-tetramethylbenzidine, and finally determined by measuring the catalysate at characteristic wavelength of 450 nm. This proposed biosensor was able to separate ∼70% of target Salmonella cells from 50 mL of bacterial sample and quantitatively detect Salmonella from 101 to 104 CFU/mL in 2.5 h with the lower detection limit of 11 CFU/mL. The mean recovery for Salmonella in spiked chicken carcass was about 109.8%. This new magnetic grid separation method was first time reported for efficient separation of target bacteria from very large volume of sample to greatly improve the sensitivity of this biosensor and could be used with various biosensing assays for practical applications in routine detection of foodborne pathogens without any bacterial pre-enrichment. • Magnetic particle chains were successfully formed in fluidic channel using iron grid. • Magnetic grid separation was very effective to separate target bacteria from large volume. • Pt@ZIF-8 with HRP-mimic enzymatic activity could greatly amplify biological signals. • This biosensor was able to detect Salmonella as low as 11 CFU/mL in 2.5 h. • This biosensor might be used for bacterial routine screening without any pre-enrichment. [ABSTRACT FROM AUTHOR]