Evaluation of a novel enzyme-linked immunosorbent assay for serological diagnosis of porcine proliferative enteropathy

Boesen, H. T.; Jensen, Tim Kåre; Möller, Kristian; Nielsen, Lisbeth Harm; Jungersen, Gregers

doi:10.1016/j.vetmic.2005.05.004

Cited by 31 publications

(28 citation statements)

References 19 publications

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“…To assess the performance of a new method, the diagnostic sensitivity (DSn) and specificity (DSp) should be determined in reference samples with known history and infection status [8,9,22]. However, these samples may not reflect the actual population for which the methods are intended and thus, the accuracy of the test might vary.…”

Section: Discussionmentioning

confidence: 99%

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Evaluation of a blocking ELISA for the detection of antibodies against Lawsonia intracellularis in pig sera

Jacobson

Wallgren

Nordengrahn³

et al. 2011

Acta Vet Scand

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BackgroundLawsonia intracellularis is a common cause of chronic diarrhoea and poor performance in young growing pigs. Diagnosis of this obligate intracellular bacterium is based on the demonstration of the microbe or microbial DNA in tissue specimens or faecal samples, or the demonstration of L. intracellularis-specific antibodies in sera. The aim of the present study was to evaluate a blocking ELISA in the detection of serum antibodies to L. intracellularis, by comparison to the previously widely used immunofluorescent antibody test (IFAT).MethodsSera were collected from 176 pigs aged 8-12 weeks originating from 24 herds with or without problems with diarrhoea and poor performance in young growing pigs. Sera were analyzed by the blocking ELISA and by IFAT. Bayesian modelling techniques were used to account for the absence of a gold standard test and the results of the blocking ELISA was modelled against the IFAT test with a "2 dependent tests, 2 populations, no gold standard" model.ResultsAt the finally selected cut-off value of percent inhibition (PI) 35, the diagnostic sensitivity of the blocking ELISA was 72% and the diagnostic specificity was 93%. The positive predictive value was 0.82 and the negative predictive value was 0.89, at the observed prevalence of 33.5%.ConclusionThe sensitivity and specificity as evaluated by Bayesian statistic techniques differed from that previously reported. Properties of diagnostic tests may well vary between countries, laboratories and among populations of animals. In the absence of a true gold standard, the importance of validating new methods by appropriate statistical methods and with respect to the target population must be emphasized.

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Section: Discussionmentioning

confidence: 99%

“…Such methods include indirect ELISAs employing whole-cell antigens [7,8], DOC -extracted antigens [9], or targeting L. intracellularis -specific antigenic epitopes such as LPS [10], LsaA [11], or the FliC protein [12]. The stated sensitivities vary from 67 to 99.5% and the specificities from 93 to 100%.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of a blocking ELISA for the detection of antibodies against Lawsonia intracellularis in pig sera

Jacobson

Wallgren

Nordengrahn³

et al. 2011

Acta Vet Scand

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“…The ISH, IHC, qPCR, and serology procedures were performed as previously described (Boye et al 1998;Boesen et al 2005;Enoe et al 2006;Jensen et al 2006;Hjulsager et al 2009;Stahl et al 2011).…”

mentioning

confidence: 99%

Demonstration of non-specific colitis and increased crypt depth in colon of weaned pigs with diarrhea

Pedersen

Kristensen

Nielsen

2012

Veterinary Quarterly

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“…Measurements of L. intracellularis -specific IgG were done by indirect ELISA using Polysorp plates (Nunc, Roskilde, Denmark) coated with L. intracellularis deoxycholate (DOC) extract as previously described [20]. For detection of specific IgA in serum or faeces extracts, Polysorp plates were coated with L. intracellularis DOC extract diluted 1:400 and with serum samples diluted 1:12.5 in blocking buffer or with undiluted faeces extracts.…”

Section: Methodsmentioning

confidence: 99%

Cell-mediated and humoral immune responses in pigs following primary and challenge-exposure to Lawsonia intracellularis

Cordes

Riber

Jensen

et al. 2012

Vet Res

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To investigate immune responses upon re-infection with Lawsonia intracellularis, local and peripheral humoral and cell-mediated immune responses to primary and challenge inoculations were studied in 22 pigs. Pigs were orally inoculated with virulent L. intracellularis at the age of 5-6 weeks, treated with antibiotics and challenged with a re-inoculation (RE) at the age of 12 weeks. Treatment control (TC) pigs received only the primary inoculation and challenge control (CC) pigs received only the secondary inoculation at 12 weeks of age. Following this regimen, all RE pigs were protected against the re-infection as defined by reduced colonisation and pathology of intestinal mucosa, absence of bacterial shedding and without increase in serum acute phase protein response. In the protected RE pigs, serum IgG responses were variable with both high and low responders. Serum IgA responses were not boosted by the re-inoculation, since identical intestinal IgA responses developed in response to the inoculation in both the susceptible CC pigs and the protected RE pigs. A memory recall cell-mediated immune response developed in RE pigs which was significantly stronger compared to the primary response in age-matched CC pigs as assessed by whole blood IFN-γ assay and by calculation of IFN-γ integrated median fluorescence intensity (iMFI) after flow cytometry. The major IFN-γ producing cells were identified as CD8+ and CD4+CD8+ double positive lymphocytes. The results indicate that cell-mediated immune responses are likely mediators of protective immunity against L. intracellularis, with CD8+ effector cells and CD4+CD8+ double positive memory T cells as main contributors to the antigen-specific IFN-γ production.

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Evaluation of a novel enzyme-linked immunosorbent assay for serological diagnosis of porcine proliferative enteropathy

Cited by 31 publications

References 19 publications

Evaluation of a blocking ELISA for the detection of antibodies against Lawsonia intracellularis in pig sera

Evaluation of a blocking ELISA for the detection of antibodies against Lawsonia intracellularis in pig sera

Demonstration of non-specific colitis and increased crypt depth in colon of weaned pigs with diarrhea

Cell-mediated and humoral immune responses in pigs following primary and challenge-exposure to Lawsonia intracellularis

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