Use of an enzyme-linked immunosorbent assay for detection of Treponema hyodysenteriae infection in swine

Wright, James C.; Wilt, G. R.; Reed, Robert; Powe, T.A.

doi:10.1128/jcm.27.3.411-416.1989

Cited by 14 publications

(8 citation statements)

References 2 publications

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“…At present, the detection and identification of S. hyodysentenae in fecal specimens from pigs with suspected swine dysentery require isolation of the organism on artificial medium and confirmation by serotyping or protein profile analysis followed by Western blotting (25,26,32). These procedures are time-consuming, labor-intensive, and costly.…”

Section: Discussionmentioning

confidence: 99%

Characterization of two DNA probes specific for Serpulina hyodysenteriae

Sotiropoulos¹,

Smith²,

Coloe³

1993

J Clin Microbiol

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Two DNA probes, one 1.1-and one 0.75-kb probe, specific for Serpulina hyodysenteriae were isolated from a genomic library generated from virulent S. hyodysenteriae 5380. These probes are highly specific and react with all S. hyodysenteriae strains tested. Under stringent conditions, the DNA probes did not react with the nonpathogenic species Serpulina innocens or with other species of enteric bacteria, including Escherichia coli. Both probes are able to detect S. hyodysenteriae in colony blot hybridizations, and when applied to fecal specimens, they can detect 104 S. hyodysenteriae cells in 0.1 g of seeded fecal matter. Both probes can detect S. hyodysenteriae in fecal specimens from swine with clinical signs of swine dysentery after experimental challenge and from swine from a herd with an acute outbreak of swine dysentery. These probes have application as a diagnostic tool in veterinary microbiology.

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Section: Discussionmentioning

confidence: 99%

Characterization of two DNA probes specific for Serpulina hyodysenteriae

Sotiropoulos¹,

Smith²,

Coloe³

1993

J Clin Microbiol

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“…This immunity appears to be antibody mediated, since colonic loops are protected against infection after passive transfer of convalescent serum antibodies (Joens et al, 1979;Harris & Glock, 1981). While B. hyodysenteriae-specific antibodies have been detected during and after swine dysentery (Joens et al, 1984(Joens et al, , 1985Rees et al, 1989;Wright et al, 1989), they have not been related to recovery from disease but instead regarded as an indication of prolonged or recent exposure to B. hyodysenteriae (Rees et al, 1989). To date, the protection induced by vaccines has only been associated with reduced severity of lesions without decreasing bacterial shedding (Waters et al, 1999a, b;Hontecillas et al, 2002) or induction of B. hyodysenteriae-specific IgA secretion (La et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Immunological alterations during the clinical and recovery phases of experimental swine dysentery

Jonasson¹,

Andersson²,

Råsbäck³

et al. 2006

Journal of Medical Microbiology

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The aim of this study was to examine changes in the systemic immune response during the incubation period and following the onset of clinical swine dysentery, including the recovery period. Ten healthy conventional pigs were inoculated with Brachyspira hyodysenteriae. Blood was sampled at pre-inoculation, at days 4 and 14 post-inoculation, during the first 4 days with clinical signs of dysentery and at days 1, 3, 7, 11 and 15 of the recovery period. Eight pigs developed haemorrhagic diarrhoea. Flow-cytometric analyses of lymphocyte subpopulations showed that all animals, including the two that remained healthy, had an increase in CD8a + CD4 " cells and cd T cells at days 4 and 14 post-inoculation. In addition, an increase in CD4 + CD8a + cells and CD8a + CD8b + cells was observed at days 4 and 14 post-inoculation in animals that developed dysentery. During clinical signs of dysentery, the acute-phase protein serum amyloid A was increased. There was a two-to threefold increase in both neutrophils and monocytes during signs of dysentery and at the beginning of the recovery period. The numbers of CD8a + CD8b " CD4 " , CD45RA " lymphocytes also increased during the dysentery period. Circulating CD21 + cells and CD21 + CD45RA " cells decreased at the end of the incubation period, during signs of dysentery and at the beginning of the recovery period. The dysentery-affected animals developed antibodies to B. hyodysenteriae-specific antigens (~16 kDa and~30 kDa) from the first day of recovery, and cd T cells showed an increase during the recovery period. In comparison with pre-inoculation, increased numbers of monocytes, neutrophils, CD8a + CD8b " CD4 " lymphocytes and CD45RA " lymphocytes were observed during clinical dysentery. Increased numbers of neutrophils, cd T cells and specific antibodies were seen during the recovery period. Taylor and Alexander (1971) showed that swine dysentery is caused by the Gram-negative spirochaete Brachyspira hyodysenteriae. The main clinical signs are severe mucohaemorrhagic diarrhoea and a reduced general appearance. The typical manifestations in the large intestine are excessive mucus production, haemorrhage and tissue necrosis. Due to the increased antimicrobial resistance of B. hyodysenteriae to the few antibiotic treatments available (Lobova et al., 2004), a better knowledge of the host responses during infection is essential for development of prophylactic measures. In pigs, macrophages and neutrophils are found in colonic lesions at later stages of swine dysentery, but the role of these cells is still unclear (Albassam et al., 1985). Experimental infections with B. hyodysenteriae in mice demonstrate that neutrophils are of great importance in mediating the severe oedema and structural lesions that occur in the intestine during the infection , and may thus be part of the pathogenesis. However, the commensal intestinal flora has also been shown to be necessary for the development of the disease (Whipp et al., 1979; Neef et al., 1994). Among lymphocyte subsets, CD8 + cells appear ...

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“…Enzyme-linked immunosorbent assays (ELISAs) have the potential to be used as rapid, sensitive and reproducible procedures for quantifying serum antibodies to B. hyodysenteriae, and providing indirect evidence of the current or past disease status of an individual pig, and/or of the herd of origin. ELISAs using sonicated whole cells or lipooligosaccharide (LOS) from B. hyodysenteriae as plate-coating antigens have been reported to be sensitive enough to allow detection of SD at a herd level, if sufficient pigs are tested (Joens et al, 1982;Wright et al, 1989;Smith et al, 1991;Song et al, 2012). Unfortunately, false-positive reactions hampered further development of these assays (La and Hampson, 2001).…”

Section: Introductionmentioning

confidence: 99%

Development of a serological ELISA using a recombinant protein to identify pig herds infected with Brachyspira hyodysenteriae

Song

Phillips

et al. 2015

The Veterinary Journal

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Development of a serological ELISA using a recombinant protein to identify pig herds infected with brachyspira hyodysenteriae, The Veterinary Journal (2015), http://dx.doi.org/doi: 10.1016/j.tvjl.2015.08.021. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.  The new test should be useful for screening herds for evidence of swine dysentery 21 Abstract 22Brachyspira hyodysenteriae is an anaerobic spirochaete that can induce swine 23 dysentery (SD), a severe mucohaemorrhagic colitis in grower and fattener pigs. The aim of this 24 study was to develop a serological ELISA for use as a screening method to detect evidence of 25 herd infection. Bioinformatic analysis of the complete genome sequence of strain WA1 was 26 used to identify genes predicted to encode outer membrane proteins. Twenty candidate genes 27 were expressed in an Escherichia coli mediated system, and purified as histidine-tagged 28 recombinant proteins. Selection of optimal antigens under different conditions was conducted 29 using Western blot and ELISA with a range of pig sera from infected and uninfected pigs. 30From this analysis, three recombinant proteins were selected as being most suitable for use as 31antigens. These antigens then were tested under optimized conditions in an indirect ELISA 32 detecting IgG2 using 1551 sera from healthy pigs at slaughter, comprising 896 from 18 herds 33 considered to be free from SD and 655 from 12 infected herds. 34 35Using a cut-off value for positivity of the mean plus five standard deviations of the 36 mean for the negative sera, the best overall results were obtained with the ELISA using antigen 37 H114, which was 100% specific and 91.7% sensitive at detecting the reported status of the 38herds. This new ELISA should be a useful adjunct for detecting and monitoring the status of 39 herds with respect to the presence of B. hyodysenteriae, and should prove useful for 40 understanding the dynamics of infection in herds where the spirochaete is present. 41

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Use of an enzyme-linked immunosorbent assay for detection of Treponema hyodysenteriae infection in swine

Cited by 14 publications

References 2 publications

Characterization of two DNA probes specific for Serpulina hyodysenteriae

Characterization of two DNA probes specific for Serpulina hyodysenteriae

Immunological alterations during the clinical and recovery phases of experimental swine dysentery

Development of a serological ELISA using a recombinant protein to identify pig herds infected with Brachyspira hyodysenteriae

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