Within-day repeatability for absolute quantification of Lawsonia intracellularis bacteria in feces from growing pigs

Ken Steen Pedersen, Klaus H. Pedersen, Charlotte Kristiane Hjulsager, Lars Erik Larsen, Marie Ståhl, Helle Stege, Øystein Angen, Jens Peter Nielsen

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    Abstract

    Absolute quantification of Lawsonia intracellularis by real-time polymerase chain reaction (PCR) is now possible on a routine basis. Poor repeatability of quantification can result in disease status misclassification of individual pigs when a single fecal sample is obtained. The objective of the current study was to investigate overall variation within a day for fecal numbers of L. intracellularis bacteria determined by real-time PCR in growing pigs. From each of 30 pigs with an infection of L. intracellularis, 5 fecal samples were collected within 1 day. A total of 150 fecal samples were obtained and subjected to quantitative PCR (qPCR) testing. Mean fecal dry matter content was 14.3% (standard deviation = 4.5%). Two pigs (6.7%) alternated between being L. intracellularis qPCR positive and negative. For 28 pigs, the excreting levels of L. intracellularis were within the dynamic range of the qPCR assay at all sampling points. For these 28 pigs, the mean excretion level of L. intracellularis was 6.1 log10 bacteria/g feces (standard deviation = 1.2 log10 bacteria/g feces). The maximum observed difference between 2 fecal samples from the same pig was 1.1 log10 bacteria/g feces. The average standard deviation for individual pigs was 0.27 log10 bacteria/g feces. The average coefficient of variation within pigs was 0.04, ranging from 0.006 to 0.08. The results imply that absolute quantification of L. intracellularis by qPCR has acceptable repeatability within 1 day. However, a population-specific proportion of pigs alternating between positive and negative test results must be expected.
    Original languageEnglish
    JournalJournal of Veterinary Diagnostic Investigation
    Volume24
    Issue number5
    Pages (from-to)968-970
    ISSN1040-6387
    DOIs
    Publication statusPublished - 2012

    Keywords

    • Fecal samples
    • Lawsonia intracellularis
    • Pigs
    • Quantitative real-time polymerase chain reaction
    • Within-day variation

    Cite this

    Pedersen, K. S., Pedersen, K. H., Hjulsager, C. K., Larsen, L. E., Ståhl, M., Stege, H., Angen, Ø., & Nielsen, J. P. (2012). Within-day repeatability for absolute quantification of Lawsonia intracellularis bacteria in feces from growing pigs. Journal of Veterinary Diagnostic Investigation, 24(5), 968-970. https://doi.org/10.1177/1040638712452111