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Performance Assessment and Validation of a Sensitive Immunoassay for Detection of Ruminant and Porcine Heat Stable Proteins in Compound Animal Feedstuffs

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Sandwich ELISAs have been produced to detect ruminant and porcine heat stable proteins which have been heated to 130°C, 2.7 bar (absolute pressure), for 30 mins and incorporated into compound animal feedstuffs (CAFs). Sample preparation involves heat treatment, removal of CAF proteins and analyte concentration. The test system incorporates a separate ELISA to detect non-specific binding of certain plant derived CAF components, each of which are identified. Both ruminant and porcine heat stable antigens are detected at a concentration of 1250 ppm. %CVs obtained within assay were 6.97% and between assay 9.07%. The maximum calculated false negative rate over 3 years of routine operation of the ELISAs was <1%. Each assay plate incorporates 12 negative controls permitting an additional 3SD calculation to overcome any in-plate variation.
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Keywords: > 130°C (2.7 bar); COMPOUND ANIMAL FEEDSTUFFS; ELISA; IMMUNOASSAY; PROTEIN REMOVAL; RUMINANT AND PORCINE PROTEINS

Document Type: Research Article

Affiliations: 1: Veterinary Laboratories Agency, Luddington Regional Centre, Stratford-upon-Avon, Warwickshire CV37 9SJ, UK 2: Veterinary Laboratories Agency, Weybridge, New Haw, Addlestone, Surrey KT15 3NB, UK

Publication date: December 1, 2000

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