Abstract
AbstractA sandwich enzyme labelled immunoassay (ELISA) has been developed for the detection of Campylobacter jejuni in food using magnetic poly(glycidyl methacrylate) (PGMA) microspheres as an alternative solid phase. The microspheres (c. 2.5 μm in diameter) were prepared by dispersion polymerization in the presence of carboxymethylcellulose-coated Fe3O4 - obtained by precipitation of Fe(II) and Fe(III) salts with aqueous ammonia - finely dispersed in ethanol/water solution using poly(vinylpyrrolidone) and 2,2’-azoisobutyronitrile as stabilizer and initiator, respectively. PGMA microspheres were ammonolyzed and the antibody against Campylobacter jejuni was immobilized via the 2,4,6-trichloro-1,3,5-triazine method. The optimal concentration of the second hen IgY was 50 μg/ml and the optimal concentration of the labelled antibody (rabbit anti-hen IgY immunoglobulin-horseradish peroxidase complex) reached 4 μg/ml. The detection limit of the assay was 4.8⋅106 cfu/ml. A limited number of artificially contaminated and uncontaminated food samples have been tested. Only one false-negative and one false-positive result (out of 18) were observed.
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