We have developed a rapid and sensitive method for immunomagnetic separation (IMS) of Salmonella along with their real time detection via PCR. Silica-coated magnetic nanoparticles were functionalized with carboxy groups to which anti-Salmonella antibody raised against heat-inactivated whole cells of Salmonella were covalently attached. The immuno-captured target cells were detected in beverages like milk and lemon juice by multiplex PCR and real time PCR with a detection limit of 104 cfu.mL-1 and 103 cfu.mL-1, respectively. We demonstrate that IMS can be used for selective concentration of target bacteria from beverages for subsequent use in PCR detection. PCR also enables differentiation of Salmonella typhi and Salmonella paratyphi A using a set of four specific primers. In addition, IMS-PCR can be used as a screening tool in the food and beverage industry for the detection of Salmonella within 3-4 h which compares favorably to the time of several days that is needed in case of conventional detection based on culture and biochemical methods.
- biofunctionalized nanoparticles
- immunomagnetic separation
- multiplex detection