Evaluation of a Novel Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against Salmonella, Employing a Stable Coating of Lipopolysaccharide-Derived Antigens Covalently Attached to Polystyrene Microwells

Author:

Wiuff Camilla1,Jauho Eva Stenbæk2,Stryhn Henrik1,Andresen Lars Ole1,Thaulov Karina2,Boas Ulrik2,Jakobsen Mogens Havsteen2,Heegaard Peter M. H.1

Affiliation:

1. Danish Veterinary Laboratory, Bülowsvej 27, DK-1790 Copenhagen V, Denmark

2. Exiqon A/S, Bygstubben 9, DK-2950 Vedbæk, Denmark

Abstract

Polysaccharides derived from Salmonella typhimurium lipopolysaccharide (LPS) representing the O-antigen factors 1, 4, 5, and 12 and the O-antigen factors 6 and 7 from Salmonella choleraesuis LPS were derivatized with the photoreactive compound anthraquinone and subsequently covalently coupled to microtiter polystyrene plates by ultraviolet irradiation. Both polysaccharide antigens could be coupled simultaneously to the same microtiter plate. The coated surface was used in indirect ELISA for the determination of serum antibodies from pigs infected with bacteria of the two Salmonella groups and from uninfected pigs. This ELISA proved itself by having a good long-term durability and a high degree of reproducibility, including low day-to-day variations and low interplate variations. Furthermore, the ELISA showed good specificity and sensitivity when data were compared with the optical density levels of a panel of pig sera as determined by a conventional ELISA on the basis of passive coating of the two Salmonella LPS antigens (the mix-ELISA). The covalent anthraquinone mix-ELISA shows promise as a stable and durable alternative to the existing conventional ELISA for serological surveillance of Salmonella infections in pigs.

Publisher

SAGE Publications

Subject

General Veterinary

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