Application and evaluation of enzyme-linked immunosorbent assay and immunoblotting for detection of antibodies to<i>Treponema hyodysenteriae</i>in swine

Smith, Stuart; Barrett, Louise M.; Muir, Tom W.; Christopher, W. L.; Coloe, Peter J.

doi:10.1017/s0950268800048937

Cited by 10 publications

(7 citation statements)

References 14 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%

“…The preferred method for characterizing S. hyodysenteriae is sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis of cellular proteins and Western blotting (immunoblotting) of the isolates with S. hyodysenteiaespecific antisera (25,26). However, restriction enzyme analysis (6), serotyping (2,20), and in vitro enteropathogenicity studies have also been used (12,15,21,28), but all these tests are time-consuming, labor-intensive, and expensive.…”

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confidence: 99%

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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%

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confidence: 99%

Characterization of two DNA probes specific for Serpulina hyodysenteriae

Sotiropoulos¹,

Smith²,

Coloe³

1993

J Clin Microbiol

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“…Swine dysentery has been detected in herds by the use of an ELISA in which serum antibodies are measured using whole sonicated cells of B. hyodysenteriae to coat the wells of the ELISA plate (Wright et al, 1989;Smith et al, 1991). This form of ELISA appears to be sensitive enough to detect the majority of infected herds.…”

Section: Enzyme-linked Immunosorbent Assay With Sonicated Bacteria Asmentioning

confidence: 99%

Serologic detection ofBrachyspira (Serpulina) hyodysenteriaeinfections

Hampson

2001

Anim. Health. Res. Rev.

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Swine dysentery (SD) caused by the intestinal spirochete Brachyspira hyodysenteriae is an economically important disease in pig-producing countries throughout the world. To date, no specific serologic assay is commercially available for the diagnosis of pigs with SD. Several serologic techniques have been identified in the past; however, these tests have all used either whole-cell proteins or lipopolysaccharide (LPS) as the antigen. Whole-cell antigens are plagued with false-positive reactions due to cross-reactivity with common proteins shared with other spirochetes. LPS antigens produce fewer false-positives; however, false-negatives may result due to LPS components being serogroup-specific. Generally, these techniques are useful for detecting infected herds, but are unreliable for the detection of individual infected pigs. In order to develop improved serologic tests it will be necessary to identify suitable diagnostic antigens, in particular immunogenic cell-surface structures which are specific to B. hyodysenteriae but common amongst different strains of the species. Recently, we identified and cloned a 30-kDa outer membrane lipoprotein (BmpB) which is specific to B. hyodysenteriae and is recognized by experimentally and naturally infected pigs. In this review we summarize the available serologic tests for SD, and speculate on the use of recombinant BmpB as an antigen for future development of an improved serologic test for SD diagnosis.

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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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Application and evaluation of enzyme-linked immunosorbent assay and immunoblotting for detection of antibodies toTreponema hyodysenteriaein swine

Cited by 10 publications

References 14 publications

Characterization of two DNA probes specific for Serpulina hyodysenteriae

Characterization of two DNA probes specific for Serpulina hyodysenteriae

Serologic detection ofBrachyspira (Serpulina) hyodysenteriaeinfections

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

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