A synthetic oligonucleotide probe and a cloned polynucleotide probe based on the yopA gene for detection and enumeration of virulent Yersinia enterocolitica

Kapperud, Georg; Dommarsnes, Kari; Skurnik, Mikael; Hornes, Erik

doi:10.1128/aem.56.1.17-23.1990

Cited by 31 publications

(18 citation statements)

References 25 publications

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“…This issue could be resolved in the near future if we succeed in obtaining natural isolates of the subspecies cremoris genotype and study their phenotypic properties in detail. Nucleic acid hybridization recently was introduced as a rapid tool for the identification of microorganisms (8,9,14). rRNAs are attractive candidates as targets for hybridization probes because of their unique organization, the presence of highly conserved and variable regions, and their presence in high copy number.…”

Section: Discussionmentioning

confidence: 99%

Development and application of oligonucleotide probes for identification of Lactococcus lactis subsp. cremoris

Salama

Sandine

Giovannoni

1991

Appl Environ Microbiol

152

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Lactococcus lactis subsp. cremoris is of considerable interest to the dairy industry, which relies upon the few available strains for the manufacture of cheddar cheese free of fermented and fruity flavors. The subspecies cremoris differs from related subspecies by the lack of a few phenotypic traits. Our purpose was to identify unique rRNA sequences that could be used to discriminate L. lactis subsp. cremoris from related subspecies. The 16S rRNAs from 13 Lactococcus strains were partially sequenced by using reverse transcriptase to identify domains unique to L. lactis subsp. cremoris. All five strains of the subspecies cremoris had a unique base sequence in a hypervariable region located 70 to 100 bases from the 5' terminus. In this region, all L. lactis subsp. lactis biovar diacetylactis strains examined had a sequence identical to that of L. lactis subsp. lactis 7962, which was different from other strains of the subspecies lactis by only one nucleotide at position 90 (Escherichia coli 16S rRNA structural model) (

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Section: Discussionmentioning

confidence: 99%

Development and application of oligonucleotide probes for identification of Lactococcus lactis subsp. cremoris

Salama

Sandine

Giovannoni

1991

Appl Environ Microbiol

152

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“…Further, differentiation based on rRNA testing is another of a growing number of clinical microbiological techniques that uses molecular biology in the detection of microbial pathogens and virulence factors. These techniques include DNA or RNA probes (8,24), plasmid analysis (38) and gene amplification using the polymerase chain reaction (21), In spite of its limited utility, this method is quick and easy to perform, especially in laboratories that use techniques in molecular biology, and the test can be completed within 60-90 min. The 10% SDS and the reaction buffer may be prepared in advance and stored almost indefinitely at room temperature, and an agarose gel takes only minuts to prepare.…”

Section: S 16smentioning

confidence: 99%

Rapid identification of Actinobacillus actinomycetemcomitans based on analysis of 23S ribosomal RNA

Preus

Sunday

Haraszthy

et al. 1992

Oral Microbiology and Immunology

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Actinobacillus actinomycetemcomitans is a key microorganism in the pathogenesis of several different forms of periodontal diseases. Identification of this bacterium from clinical specimens may often be complicated by the fact that the colony morphology on TSBV selective medium closely resembles that of Haemophilus aphrophilus and a key differentiating characteristic, catalase reaction, may be variable. Recent genetic studies have shown that the 23S ribosomal RNA molecule is split into two smaller forms in A. actinomycetemcomitans, but is intact in H. aphrophilus. Based on this finding, we describe a new, rapid method for identifying A. actinomycetemcomitans in which single colonies isolated from culture on TSBV agar in 5% CO2 in air are lysed, electrophoresed on 1.5% submarine agarose gels and visualized by staining with ethidium bromide. Using this assay, A. actinomycetemcomitans can be easily distinguished from morphologically similar colonies such as H. aphrophilus strains by differences in 23S rRNA within 2 h.

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“…Research on the genetic basis of the phenotypic differences between Y. enterocolitica serotypes has resulted in the discovery of a virulence-associated plasmid Gemski et al 1980) and the presence or absence of this plasmid has been used to discriminate between virulent and non-virulent strains (Bolin et al 1982;Hill et al 1983;Jagow and Hill 1986;Miliotis et al 1989;Kapperud et al 1990;Nesbakken et al 1991). Methods based on the detection of the virulence plasmid, however, may lead to false negative results, because the plasmid is easily lost during repeated subculturing of the bacteria (Zink et al 1980;.…”

Section: Introductionmentioning

confidence: 99%

Digoxigenin‐labelled inv‐ and ail‐probes for the detection and identification of pathogenic Yersinia enterocolitica in clinical specimens and naturally contaminated pig samples

Goverde

Jansen

Brunings

et al. 1993

Journal of Applied Bacteriology

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A non-radioactive colony hybridization method was developed for the rapid detection of Yersinia enterocolitica in primary isolates and for differentiation between pathogenic and non-pathogenic strains. The method is based on, respectively, the presence of the inv-locus in all Yersinia spp. and the presence of the ail-gene in pathogenic Y. enterocolitica only. Hybridization results with ail-probes of 132 strains of Y. enterocolitica were in good agreement with pathogenicity phenotypes as indicated by a tissue culture invasion (TCI) assay and by serotyping. All TCI+ strains and only two TCI- strains were positive by hybridization with ail. Hybridization results with inv- or ail-probes of 150 primary isolates of human, animal or slaughterhouse origin were compared with those of conventional methods to detect and identify Y. enterocolitica. All samples that were positive for Yersinia spp. by cultivation (four of 66) or were positive for pathogenic Y. enterocolitica by cultivation and serotyping (six of 84) were also positive by hybridization with, respectively, the inv- or ail-probe. In three slaughterhouse swab samples, in which Yersinia spp. were not detected by cultivation (2%), strong positive hybridization signals were obtained with the inv- and/or ail-probe. Four other swab samples which were negative by cultivation produced weak positive signals by hybridization with inv- and/or ail-probes. These results indicate that the method can be used for (1) the identification of pathogenic Y. enterocolitica isolates and (2) the detection of Yersinia spp. in primary isolates of naturally contaminated samples.

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A synthetic oligonucleotide probe and a cloned polynucleotide probe based on the yopA gene for detection and enumeration of virulent Yersinia enterocolitica

Cited by 31 publications

References 25 publications

Development and application of oligonucleotide probes for identification of Lactococcus lactis subsp. cremoris

Development and application of oligonucleotide probes for identification of Lactococcus lactis subsp. cremoris

Rapid identification of Actinobacillus actinomycetemcomitans based on analysis of 23S ribosomal RNA

Digoxigenin‐labelled inv‐ and ail‐probes for the detection and identification of pathogenic Yersinia enterocolitica in clinical specimens and naturally contaminated pig samples

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