Separation of Listeria from cheese and enrichment media using antibody-coated microbeads and centrifugation.

An immunoseparation system for the separation of Listeria from enriched cheese samples was developed. The system utilizes polystyrene microbeads (3.8 microm in diameter) coated with covalently bound anti-Listeria genus-specific antibody. The beads were incubated with cheese enriched in half-Fraser broth and the bead-bacterial complex was separated by centrifugation at 110 g then spread on selective agar plates. Although cross-reactivity with certain Gram-positive bacteria (Staphylococcus saprophyticus and Arthrobacter sp.) was determined, this had no negative effect on capture effectiveness of the beads to Listeria spp. The minimum density of Listeria cells positively detected by immunoseparation with subsequent plating was 10(0) cfu/ml. The application of the separation method facilitates a reduction in the time of Listeria detection in cheese by 2 days without affecting the sensitivity.