Simultaneous detection of antibodies to five Actinobacillus pleuropneumoniae serovars using bead-based multiplex analysis

Sanne Schou Berger, Klara Tølbøl Lauritsen, Ulrik Boas, Peter Lind, Lars Ole Andresen

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    We have developed and made a preliminary validation of a bead-based multiplexed immunoassay for simultaneous detection of porcine serum antibodies to Actinobacillus pleuropneumoniae serovars 1, 2, 6, 7, and 12. Magnetic fluorescent beads were coupled with A. pleuropneumoniae antigens and tested with a panel of serum samples from experimentally infected pigs and with serum samples from uninfected and naturally infected pigs. The multiplex assay was compared to in-house ELISAs and complement fixation (CF) tests, which have been used for decades as tools for herd classification in the Danish Specific Pathogen Free system. Assay specificities and sensitivities as well as the corresponding cutoff values were determined using receiver operating characteristic (ROC) curve analysis, and the A. pleuropneumoniae multiplex assay showed good correlation with the in-house ELISAs and CF tests with areas under ROC curves ≥ 0.988. Benefits of multiplexed assays compared to ELISAs and CF tests include reduced serum sample volumes needed for analysis, less labor, and shorter assay time.
    Original languageEnglish
    JournalJournal of Veterinary Diagnostic Investigation
    Volume29
    Issue number6
    Pages (from-to)797-804
    ISSN1040-6387
    DOIs
    Publication statusPublished - 2017

    Keywords

    • Actinobacillus pleuropneumoniae
    • magnetic beads
    • multiplex analysis
    • swine

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