<i>Bilophila wadsworthia</i>Clinical Isolates Compared by Polymerase Chain Reaction Fingerprinting

Gerardo, S. Hunt; Marina, M.; Citron, Diane M.; Claros, Marina C.; Hudspeth, Marie K.; Goldstein, Ellie J. C.

doi:10.1086/516229

Cited by 8 publications

(6 citation statements)

References 8 publications

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“…septica (22), would present with the same polymorphic genomic fingerprint. If anything, when using the M13 core primer, one finds different, distinct PCR fingerprint patterns within a species, as was seen in this and other studies (1,11,16,18,31). Furthermore, in the study in which Mutters et al (22) separated the three P. multocida subspecies based on DNA-DNA homology, only 4 dulcitol-negative, sorbitol-negative isolates (P. multocida subsp.…”

Section: Discussionsupporting

confidence: 58%

Pasteurella multocida subsp. multocida and P. multocida subsp. septica Differentiation by PCR Fingerprinting and α-Glucosidase Activity

et al. 2001

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Pasteurella multocida is composed of three subspecies that are often differentiated by fermentation of sorbitol and dulcitol. We studied 35 dulcitol-negative P. multocida isolates from infected dog and cat bite wounds, 16 of which yielded weak and/or conflicting fermentation reactions in Andrades sorbitol, thus making it difficult to distinguish between the two dulcitol-negative subspecies of P. multocida, i.e., P. multocida subsp. multocida and P. multocida subsp. septica. All isolates and two control strains were further analyzed using a PCR fingerprinting technique with a single primer (M13 core) and assessed for ␣-glucosidase (␣-Glu) activity. Although the PCR fingerprint patterns and ␣-Glu activity did not correlate well with the sorbitol fermentation reactions, they did correlate well with each other. All strains identified as P. multocida subsp. septica were positive for ␣-Glu activity and exhibited the group I PCR fingerprint profile. All strains categorized as P. multocida subsp. multocida displayed either the group II or group III PCR fingerprint profile; 9 of 11 of these isolates were ␣-Glu negative. These data suggest that both PCR fingerprinting and ␣-Glu activity provide reliable means for differentiating P. multocida subsp. multocida from P. multocida subsp. septica, particularly in strains that produce weak and/or discrepant sorbitol fermentation reactions.

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Section: Discussionsupporting

confidence: 58%

Pasteurella multocida subsp. multocida and P. multocida subsp. septica Differentiation by PCR Fingerprinting and α-Glucosidase Activity

et al. 2001

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“…Convex colonies with black centres and translucent margins were observed in each culture, which indicated the presence of B. wadsworthia (Baron et al, 1989). These identifications were confirmed by PCR fingerprinting using the primer T3B (Hunt Gerardo et al, 1997), yielding in all cases the characteristic band for B. wadsworthia.…”

Section: Enrichment Of B Wadsworthia From Animal Faecesmentioning

confidence: 60%

“…After about four subcultures to fresh liquid medium, 11 enrichments were considered stable and positive. These identifications were confirmed by PCR fingerprinting using the primer T3B (Hunt Gerardo et al, 1997), yielding in all cases the characteristic band for B. wadsworthia. the morphotype of B. wadsworthia (Baron et al, 1989).…”

Section: Enrichment Of B Wadsworthia From Animal Faecesmentioning

confidence: 66%

“…PCR products and a 1 kb DNA ladder molecular size marker (MBI Fermentas) were loaded on to a 1.5% agarose gel, which was subsequently stained with ethidium bromide (Sambrook et al, 1989), to evaluate the PCR. To verify the identity of candidate B. wadsworthia isolates obtained from the enrichment cultures, the PCR fingerprinting technique was used (Hunt Gerardo et al, 1997). Single primer amplifications were carried out with the tDNA primer T3B, and PCR products were visualized on agarose gels.…”

Section: Molecular Methodsmentioning

confidence: 99%

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Identification ofBilophila wadsworthiaby specific PCR which targets the taurine:pyruvate aminotransferase gene

Laue

Smits

Schumacher

et al. 2006

FEMS Microbiology Letters

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The bile-resistant, strictly anaerobic bacterium Bilophila wadsworthia is found in human faecal flora, in human infections and in environmental samples. A specific PCR primer set for the gene encoding the first metabolic enzyme in the degradative pathway for taurine in B. wadsworthia, taurine:pyruvate aminotransferase (tpa), was developed and tested. In addition, enrichment cultures were started from faecal samples of primates and felines and shown to contain B. wadsworthia. These were subcultured on agar media and then identified by PCR fingerprinting. PCR for tpa was successful in all positive enrichment cultures and showed no amplification signal in a variety of other bacterial species. Therefore, this PCR method could be a promising tool for rapid detection of B. wadsworthia in biological samples.

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“…Endotoxic and procoagulant activities have been identified in B. wadsworthia (15), and an in vitro study suggested that it may be able to attach to epithelial cells of the colon (8). Separate subgroups or strains of B. wadsworthia have been indicated by DNA fingerprinting studies (9). The possible isolation of B. wadsworthia from healthy or diseased nonhuman hosts has not previously been reported.…”

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confidence: 99%

Colonic Infection by Bilophila wadsworthia in Pigs

et al. 2001

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Bilophila wadsworthia is a common inhabitant of the human colon and has been associated with appendicitis and other local sites of inflammation in humans. Challenge-exposure or prevalence studies in laboratory and other animals have not been reported. B. wadsworthia is closely related phylogenetically to Desulfovibrio sp. and Lawsonia intracellularis, which are considered colon pathogens. We developed a PCR specific for B. wadsworthia DNA. Samples of bacterial DNA extracted from the feces of pigs on six farms in Australia and four farms in Venezuela were examined. Specific DNA of B. wadsworthia was detected in the feces of 58 of 161 Australian and 2 of 45 Venezuelan pigs, results comprising 100% of the neonatal pigs, 15% of the weaned grower pigs, and 27% of the adult sows tested. Single-stranded conformational polymorphism analysis of PCR product DNA derived from pigs or from known human strains showed an identical pattern. Histologic examination of the intestines of weaned B. wadsworthia-positive pigs found no or minor specific lesions in the small and large intestines, respectively. B. wadsworthia is apparently a common infection in neonatal pigs, but its prevalence decreases after weaning. The possible role of B. wadsworthia as an infection in animals and in human colons requires further study.

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Bilophila wadsworthiaClinical Isolates Compared by Polymerase Chain Reaction Fingerprinting

Cited by 8 publications

References 8 publications

Pasteurella multocida subsp. multocida and P. multocida subsp. septica Differentiation by PCR Fingerprinting and α-Glucosidase Activity

Pasteurella multocida subsp. multocida and P. multocida subsp. septica Differentiation by PCR Fingerprinting and α-Glucosidase Activity

Identification ofBilophila wadsworthiaby specific PCR which targets the taurine:pyruvate aminotransferase gene

Colonic Infection by Bilophila wadsworthia in Pigs

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