Evaluation of a Commercial ELISA for Detection of Ruminant Processed Animal Proteins in Non-Ruminant Processed Animal Proteins

M.G.E.G. Bremer, R.J.C.F. Margry, J.C.H. Vaessen, A.M.H. van Doremalen, J.G.P. van der Palen, R.G.C. van Kaathoven, A.E.M. Kemmers-Voncken, L.W.D. van Raamsdonk

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    9 Citations (Scopus)


    Due to a growing aquaculture industry, demand for high-quality proteins for aquatic feeds is increasing. Non-ruminant processed animal proteins (PAPs) have shown great potential for this purpose. Safe reintroduction of non-ruminant PAPs in aqua feed requires methods that can discriminate ruminant and non-ruminant PAPs at contamination levels at or below 2%. Because the official European Union method lacks species specificity, the performance of MELISA-TEK™ Ruminant, a commercial immunoassay, combined with the MELISA-TEK High Sensitivity Sample Extraction kit was evaluated. Various non-ruminant PAPs spiked with ruminant PAPs (processed at 133, 137, 141, and 145°C) were analyzed. Results showed an overall specificity of 99%, indicating no cross-reaction with non-ruminant PAPs. The sensitivity of the assay strongly depended on both processing temperature and proportion of muscle fibers of the ruminant PAPs. Overall sensitivity of samples with 1 and 2% ruminant PAPs was 92 and 100%, respectively. For ruminant PAPs processed at 133 and 137°C, the sensitivity was 100% for both 1 and 2% ruminant spikes. Overall accuracies were 96 and 99% for 1 and 2% ruminant spikes, respectively. In conclusion, the MELISA-TEK Ruminant assay showed satisfactory results, which makes it a suitable candidate method to enable safe reintroduction of non-ruminant PAPs in aqua feed.
    Original languageEnglish
    Pages (from-to)552-559
    JournalJournal of AOAC International
    Issue number3
    Publication statusPublished - 2013


    • plant sterilization conditions
    • shrimp litopenaeus-vannamei
    • linked-immunosorbent-assay
    • bone meals
    • classical microscopy
    • rendered meat
    • by-products
    • feed
    • pcr
    • identification


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