RAPD fingerprinting for the distinction ofPrevotella intermediasensu stricto fromPrevotella nigrescens

Paquet, Caroline; Mouton, Christian

doi:10.1006/anae.1997.0077

Cited by 11 publications

(13 citation statements)

References 35 publications

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“…Comparisons between the fingerprints were performed as described previously [18]. Briefly, computer-assisted analysis of the scanned fingerprint was performed with the Taxotron1 package, a set of programs for molecular systematics [19].…”

Section: Rep-pcr Products Analysismentioning

confidence: 99%

Use of rep-PCR to define genetic relatedness among Bacteroides fragilis strains

Moraes¹,

Gonçalves

Mouton

et al. 2000

Journal of Medical Microbiology

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Bacteroides fragilis, a component of the normal flora and an important anaerobic pathogen in non-intestinal endogenous infections, has recently been associated with enteric diseases. In this study, 41 B. fragilis strains were analysed in relation to their genetic diversity. This collection included two reference strains (ATCC 23745 and 25285), 20 isolates from non-intestinal infections, six from intestinal infections, five from intestinal microflora and eight from an aquatic environment. The fingerprints were generated by using two repetitive sequences (REP and ERIC) as primers to PCR (rep-PCR). A dendrogram was obtained with the Taxotron1 Program. Three clusters (threshold genotypes I, II and III) were observed when the genetic distance was 0.30. These results confirm previous data found regarding the genotypical diversity of B. fragilis.

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Section: Rep-pcr Products Analysismentioning

confidence: 99%

Use of rep-PCR to define genetic relatedness among Bacteroides fragilis strains

Moraes¹,

Gonçalves

Mouton

et al. 2000

Journal of Medical Microbiology

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“…Enzyme electrophoresis of malate dehydrogenase was performed on non-denaturing discontinuous polyacrylamide gel as described elsewhere (23). The migration of malate dehydrogenase of each study strain was determined using the type strains P. intermedia ATCC 2561 1 and P. nigrescens NCTC 9336 as mobility references on each gel.…”

Section: Electrophoresis Of the Enzyme Malate Dehydrogenasementioning

confidence: 99%

“…These data contrast with the report by Moore et al (19) in which homology group VPI 4197, now P. intermedia sensu stricto, was preferentially associated with healthy sites or shallow pockets, whereas an association between homology group VPI 8944, now P. nigrescens, and active, deeper periodontitis sites was observed. Furthermore, in a recent study (23) we observed that strains of P. intermedia sensu stricto as well as of P. nigrescens can be isolated from a variety of clinical situations, including gingival health, gingivitis, periodontitis and endodontic infections. Similarly, Matt6 et al (18) have reported a high occurrence of both species in periodontitis pockets.…”

mentioning

confidence: 95%

β‐lactamase‐producing strains in the species Prevotella intermedia and Prevotella nigrescens

Bernal

Guillot

Paquet

et al. 1998

Oral Microbiology and Immunology

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A total of 96 strains were collected that included laboratory strains and clinical isolates classified Prevotella intermedia sensu lato and the type strains of the species P. intermedia sensu stricto and Prevotella nigrescens. Susceptibility to amoxicillin and amoxicillin-clavulanic acid was determined by the Etest. PCR-DNA probe assays were used to speciate each strain as P. intermedia sensu stricto or P. nigrescens. By Etest, 71 strains (74%) were susceptible to both amoxicillin and amoxicillin-clavulanic acid with minimum inhibitory concentrations in the 0.016-0.064 microgram/ml range. In contrast, amoxicillin minimum inhibitory concentrations of 25 strains (26%) were in the range of 1.5-96 micrograms/ml with concomitant amoxicillin-clavulanic acid minimum inhibitory concentrations in the low range 0.016-0.38 microgram/ml, indicating a production of beta-lactamase as confirmed by nitrocefin tests. Of these beta-lactamase-producing strains, 20% (5/25) were identified as P. intermedia sensu stricto by the PCR-DNA probe assay and 72% (18/25) as P. nigrescens. Our results provide support for the major role of P. nigrescens in the failure of therapy using beta-lactam antibiotics.

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“…Both species share many phenotypic traits (20), making speciation difficult to perform. To separate the two species, one must resort to one of the following tests or techniques: DNA-DNA hybridization (17), serotyping with monoclonal antibodies (5,12), enzyme electrophoresis of malate and glutamate dehydrogenases (31), sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of soluble cellular proteins (10), restriction fragment length polymorphism analysis (8), hybridizations using 16S rRNA probes (6,32), ribotyping (4,28), or randomly amplified polymorphic DNA (RAPD) fingerprinting (22,27).…”

mentioning

confidence: 99%

“…A total of 50 strains (Table 1) were used in the present study and included as references the type strains ATCC 25611 (VPI 4197) of the species P. intermedia sensu stricto and NCTC 9336 (ϭ ATCC 33563, ϭ VPI 8944 [31]) of the species P. nigrescens. The laboratory strains and clinical isolates, which were of diverse clinical and geographical origin, were classified as P. intermedia sensu lato in our laboratory (27) by using a short set of presumptive tests and those of the Rapid ID32 A system (bioMérieux, Marcy l'Étoile, France): colonies are brown to black on enriched blood agar and show a red fluorescence under long-wave UV light; they are indole positive and do not produce catalase; and they are ␣-glucosidase positive and trypsin negative. The strains were grown to mid-log phase in Todd-Hewitt broth (BBL Microbiology Systems, Cockeysville, Md.)…”

mentioning

confidence: 99%

PCR-DNA probe assays for identification and detection of Prevotella intermedia sensu stricto and Prevotella nigrescens

Guillot

Mouton

1997

J Clin Microbiol

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The purpose of this study was to construct PCR-DNA probe assays specific for Prevotella intermedia sensu stricto and Prevotella nigrescens based on the ability of randomly amplified polymorphic DNA (RAPD) fingerprinting to generate species-specific markers. The strategy included four steps: (i) construction of firstgeneration DNA probes from a 850-bp RAPD marker for P. intermedia sensu stricto and a 1,300-bp RAPD marker for P. nigrescens, (ii) cloning and sequencing of each RAPD marker, (iii) designing of primer pairs flanking specific internal sequences of 754 bp for P. intermedia sensu stricto and of ca. 1,100 bp for P. nigrescens, and (iv) synthesis (by PCR amplification) and digoxigenin labeling of quantities of DNA probes 754 and ca. 1,100 bp in size. The PCR-DNA probe assays combine either PCR amplification of a 754-bp specific sequence in the genomic DNA of strains of P. intermedia sensu stricto and hybridization with the 754-bp digoxigeninlabeled probe or amplification of a ca. 1,100-bp sequence of P. nigrescens and hybridization with the ca. 1,100-bp probe. Specific hybridization was observed with the amplified DNAs from 25 strains of P. intermedia and 24 strains of P. nigrescens, and no reaction was observed with the PCR products from 20 foreign species. The PCR-DNA probe assays described here should allow a highly specific and sensitive detection of P. intermedia sensu stricto and P. nigrescens in mixed infections.

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RAPD fingerprinting for the distinction ofPrevotella intermediasensu stricto fromPrevotella nigrescens

Cited by 11 publications

References 35 publications

Use of rep-PCR to define genetic relatedness among Bacteroides fragilis strains

Use of rep-PCR to define genetic relatedness among Bacteroides fragilis strains

β‐lactamase‐producing strains in the species Prevotella intermedia and Prevotella nigrescens

PCR-DNA probe assays for identification and detection of Prevotella intermedia sensu stricto and Prevotella nigrescens

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