Evaluation of the An-Ident system and an indole spot test for the rapid differentiation of porcine treponemes

Bélanger, Myriam; Jacques, Mario

doi:10.1128/jcm.29.8.1727-1729.1991

Cited by 16 publications

(8 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several biochemical tests for rapid differentiation of enteropathogenic and nonpathogenic intestinal spirochetes of swine have been proposed (1,2,12,32). Although these biochemical characteristics are highly conserved among field isolates of S. hyodysenteriae, WBHIS have been shown to yield highly variable results (1-3, 17, 23, 29, 30, 39).…”

Section: Discussionmentioning

confidence: 99%

Rapid detection of Serpulina hyodysenteriae in diagnostic specimens by PCR

et al. 1994

View full text Add to dashboard Cite

PCR assay for the detection of Serpulina hyodysenteriae in diagnostic specimens was developed on the basis of sequence analysis of a recombinant clone designated pRED3C6. Clone pRED3C6, which contained a 2.3-kb DNA fragment unique to S. hyodysenteriae, was identified by screening a plasmid library of S. hyodysenteriae isolate B204 genomic DNA in Escherichia coli by colony immunoblot with the mouse monoclonal antibody 10G6/G10, which was produced against cell-free supernatant antigens from the same isolate. Southern blot analysis of Hindlll-digested genomic DNA of S. hyodysenteriae serotypes 1 through 7 and of four weakly beta-hemolytic intestinal spirochetes, including Serpulina innocens, with the 23-kb DNA fragment of pRED3C6 indicated that the cloned sequence was present exclusively in the seven serotypes of S. hyodysenteriae. An oligonucleotide primer pair for PCR amplification of a 1.55-kb fragment and an internal oligonucleotide probe were designed and synthesized on the basis of sequence analysis of the 23-kb DNA fragment of pRED3C6. Purified genomic DNAs from reference isolates of S. hyodysenteriae serotypes 1 through 9, S. innocens, weakly beta-hemolytic intestinal spirochetes belonging to genotypic groups distinct from those of reference Serpulina spp., other cultivable reference isolates of the order Spirochaetales, and enteric bacteria including Escherichia coli, Salmonella spp., Campylobacter spp., and Bacteroides vulgatus were amplified with the oligonucleotide primer pair in a hot-start PCR. The 1.55-kb products were obtained only in the presence of genomic DNA from each of the nine serotypes of S. hyodysenteriae. The specificity of the 1.55-kb products for S. hyodysenteriae was confirmed on the basis of production of a restriction endonuclease pattern of the PCR products identical to the predicted restriction map analysis of pRED3C6 and positive hybridization signal with the S. hyodysenteriae-specific internal oligonucleotide probe. By using total DNA obtained from normal swine feces inoculated with decreasing concentrations of S. hyodysenterae

show abstract

Section: Discussionmentioning

confidence: 99%

Rapid detection of Serpulina hyodysenteriae in diagnostic specimens by PCR

et al. 1994

View full text Add to dashboard Cite

show abstract

“…These methods are mainly based on enzymatic profiles of the organisms. Although these methods are of some use in discriminating between S. hyodysentenae and the apathogenic swine spirochete Serpulina innocens (1,5,23,29), often these tests are not sensitive enough to discriminate among individual isolates of S. hyodysenteriae, much less to distinguish them from other intestinal spirochetes. The outcome of these tests can also be influenced by environmental and culture conditions.…”

mentioning

confidence: 99%

Genetic similarity of intestinal spirochetes from humans and various animal species

et al. 1993

View full text Add to dashboard Cite

The chromosomal DNA of spirochetes isolated from human, swine, dog, mouse, rat, and chicken intestine or feces was subjected to restriction enzyme analysis and hybridization with three different DNA probes, derived from a flagellin gene, a hemolysin gene, and the 16S rDNA sequence of the pathogenic swine intestinal spirochete Serpulina hyodysenteriae. This genetic analysis showed that intestinal spirochetes represent a heterogenous but related population of bacteria. In general, unique genotypes were distinguished among isolates from the same host species; they were not present among isolates from other host species. This suggests the host specificity of some strains. An exception to this are isolates from humans and dogs suffering from gastrointestinal disorders; these isolates showed highly similar or even identical genotypes. None of them resembled any of the genotypes of isolates found in other host species without apparent disease.

show abstract

“…Bacteria and growth conditions. S. hyodysenteriae reference strains representing serotypes 1 (B234), 2 (B204), 3 (B169), and 4 (A-1) were provided by L. A. Joens, Department of Veterinary Science, University of Arizona, Tucson. S. hyodysenteriae serotypes 5 (B8044), 6 (B6933), and 7 (Ack 300/8) and S. innocens B256 were obtained from M. J.…”

Section: Methodsmentioning

confidence: 99%

“…S. hyodysenteriae serotypes 8 (FM 88-90) and 9 (FMV 89-3323) were from our collection (23). Nineteen isolates of S. hyodysenteriae and 11 isolates of S. innocens (3,23) were originally obtained from S. Messier, Agriculture Canada, Saint-Hyacinthe, Quebec, Canada. Bacteria were grown on solid medium by using blood agar base no.…”

Section: Methodsmentioning

confidence: 99%

Molecular characterization of Serpulina (Treponema) hyodysenteriae isolates representing serotypes 8 and 9

Jensen

Bélanger

et al. 1992

J Clin Microbiol

Self Cite

View full text Add to dashboard Cite

The study described here was carried out to further characterize reference strains of Serpulina (Treponema) hyodysenteriae representing serotypes 8 and 9. Results obtained from restriction fragment length polymorphism analysis, enteropathogenicity testing, and endotoxin profiles confirmed their identifications. Electron microscopy indicated that both strains were covered with a thin layer of capsule-like material. Immunoblot analysis indicated that an antigen in the 19-kDa region of proteinase K-digested whole cells reacted only with homologous antisemm. The serotype-specific antigens were sensitive to periodate oxidation but resistant to proteinase K digestion and migrated in the same region as purified lipopolysaccharides. Immunoblotting with proteinase K-digested whole cells appeared as useful as immunodiffusion with extracted lipopolysaccharide for the serological classification of S. hyodysenteriae. Immunogold labeling of whole cells and purified periplasmic flagella showed strong cross-reactions between S. hyodysenteriae and Serpulina innocens. Outer membrane preparations of strains representing serotypes 8 and 9 contained four major proteins which reacted with antisera against both species, and one major protein with a molecular mass of 46 kDa which reacted only with antisera against S. hyodysenteriae, irrespective of the serotype. Our findings suggest that periplasmic flagella and some outer membrane proteins are antigens common to both S. hyodysenteriae and S. innocens, whereas a 46-kDa outer membrane protein may be a species-specific antigen of S. hyodysenteriae. Finally, we propose immunoblotting as an alternative method to immunodiffusion for the serotyping of S. hyodysenteriae. Serpulina (Treponema) hyodysenteriae, a gram-negative anaerobic spirochete, is the causative agent of swine dysentery (9, 34, 35). The virulence factors of this organism have not been fully defined, but lipopolysaccharide (LPS)-endotoxin and hemolysin may be important in pathogenesis (5, 6, 24, 27). The most distinctive characteristics of S. hyodysenteriae are its strong beta-hemolytic activity and its enteropathogenicity in swine (20). Serpulina innocens (19) is morphologically identical to S. hyodysenteriae (11, 19), but it is weakly beta-hemolytic and is nonenteropathogenic for swine (19), and it shares only about 40% DNA sequence homology with S. hyodysenteriae (35). We have previously described the use of hemolysis and the ring phenomenon test in conjunction with an indole spot test for differentiation of the two species (3). Recently, oligodeoxynucleotide probes complementary to a unique region of S. hyodysenteriae 16S rRNA have been developed and used for the identification of this spirochete (14). S. hyodysenteriae LPS is responsible for serological specificity (25, 33). LPS may also play a role in protection against S. hyodysenteriae infection since protection has been shown to be serotype specific (17). Seven serotypes of S. hyodysenteriae have been described on the basis of agar gel double immunodiffusion precipitat...

show abstract

Evaluation of the An-Ident system and an indole spot test for the rapid differentiation of porcine treponemes

Cited by 16 publications

References 18 publications

Rapid detection of Serpulina hyodysenteriae in diagnostic specimens by PCR

Rapid detection of Serpulina hyodysenteriae in diagnostic specimens by PCR

Genetic similarity of intestinal spirochetes from humans and various animal species

Molecular characterization of Serpulina (Treponema) hyodysenteriae isolates representing serotypes 8 and 9

Contact Info

Product

Resources

About