Application of whole-cell DNA restriction endonuclease profiles to the epidemiology of Clostridium difficile-induced diarrhea

Kuijper, Ed J.; Oudbier, J. H.; Stuifbergen, W. N. H. M.; Jansz, A; Zanen, H. C.

doi:10.1128/jcm.25.4.751-753.1987

Cited by 88 publications

(24 citation statements)

References 19 publications

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“…Different methods have been developed to study the epidemiology of C. difficile or to identify or type strains. Analysis of restriction patterns of DNA of clinical isolates has been used for investigations of epidemiology and typing of C. difficileassociated diarrhea (24). Pantosti et al (37) used the electrophoretic patterns of extracted proteins to characterize C. difficile strains from various sources and showed correlation between certain electrophoretic patterns and virulence.…”

Section: Discussionmentioning

confidence: 99%

Phenotypic and Genotypic Diversity of the Flagellin Gene (fliC) among Clostridium difficile Isolates from Different Serogroups

Tasteyre

Karjalainen

Avesani

et al. 2000

Journal of Clinical Microbiology

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Phenotypic and genotypic diversity of the flagellin gene (fliC) of Clostridium difficile was studied in 47 isolates from various origins belonging to the serogroups A, and S3. Electron microscopy revealed 17 nonflagellated strains and 30 flagellated strains. PCR and reverse transcription-PCR demonstrated that the flagellin gene was present in all strains and that the fliC gene was expressed in both flagellated and nonflagellated strains. Southern blotting showed the presence of only one copy of the gene and three different hybridization patterns. DNA sequence analysis of fliC from the strains belonging to serogroups C, D, and X, representative of each profile, disclosed great variability in the central domain, whereas the N-and C-terminal domains were conserved. The variability of the flagellin gene fliC was further studied in the isolates by PCR-restriction fragment length polymorphism (RFLP) analysis. Nine different RFLP groups were identified (I to IX), among which three (I, VII, and VIII) corresponded to numerous serogroups whereas the six others (II, III, IV, V, VI, and IX) belonged to a single serogroup. Flagellin gene RFLP analysis could constitute an additional typing method employable in conjunction with other typing methods currently available.

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Section: Discussionmentioning

confidence: 99%

Phenotypic and Genotypic Diversity of the Flagellin Gene (fliC) among Clostridium difficile Isolates from Different Serogroups

Tasteyre

Karjalainen

Avesani

et al. 2000

Journal of Clinical Microbiology

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“…This bacterium is considered to be the main agent of nosocomially acquired diarrhea among adults [2]. Many various typing methods have been developed to investigate nosocomial outbreaks of C. di¤cile, some based on phenotype (lysotyping [3], serogroup-ing [4], SDS-PAGE [5], immunoblotting [6]) and others on genotype (plasmids [7], REA [8], ribotyping [9], PFGE [10], RAPD [11]). Recently, Kostman et al developed a method of PCR-ribotyping that was shown to be reproducible, easy to perform and cost-e¡ective [12].…”

Section: Introductionmentioning

confidence: 99%

Development of a new PCR-ribotyping method for Clostridium difficile based on ribosomal RNA gene sequencing

Bidet

1999

FEMS Microbiology Letters

101

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PCR-ribotyping, a typing method based on polymorphism in the 16S-23S intergenic spacer region, has been recently used to investigate outbreaks due to Clostridium difficile. However, this method generates bands of high and close molecular masses which are difficult to separate on agarose gel electrophoresis. To improve reading of banding patterns of PCR-ribotyping applied to C. difficile, a partial sequencing of the rRNA genes (16S and 23S) and intergenic spacer region has been performed, then a new set of primers located closer to the intergenic spacer region has been defined. The new PCR gave reproducible patterns of bands easy to separate on agarose gel electrophoresis. Each of the 10 serogroups and 11 subgroups of serogroup A produced a different pattern. This typing method has evidenced major qualities such as easiness, rapidity and reproducibility. However, its discriminatory power has to be evaluated to validate its importance as a typing tool for C. difficile. z

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“…Numerous studies have demonstrated the utility of DNA fingerprinting for epidemiological analysis of phenotypically indistinguishable microbial pathogens (1,4,15,18,19,22,36,(38)(39)(40). Ribotyping, based on restriction fragment length polymorphisms in the chromosomal DNA containing rRNA genes, has also been used as a means of detecting interspecies and interstrain differences (13,14,24).…”

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confidence: 99%

Comparison of methods for discrimination between strains of Listeria monocytogenes from epidemiological surveys

Baloga

Harlander

1991

Appl Environ Microbiol

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Total cellular DNA from 28 strains of Listeria monocytogenes isolated from food implicated in food-borne illness and from patients with listeriosis was digested with the restriction endonucleases HindIII, HaeIII, and EcoRI. Following agarose gel electrophoresis, the fragments were subjected to Southern blot hybridization with a digoxigenin-labeled cDNA probe transcribed from Escherichia coli 16S and 23S rRNA. The patterns of bands from genomic (DNA fingerprints) and rDNA fingerprints (ribotypes) were used for classifying L. monocytogenes strains, and the resulting subtypes were compared with serotyping and multilocus enzyme electrophoresis classffication schemes. A total of 15 distinct and identical groups were obtained when genomic DNA was digested with either HindIII or HaeIII. The most discriminating enzyme for ribotyping of strains was EcoRI, which divided the 28 strains of L. monocytogenes into 6 ribotype groups. DNA fingerprinting and ribotyping differentiated L. monocytogenes from other Listeria spp., including L. ivanovii, L. welshimeri, and L. innocua as well as the lactic acid bacteria Lactococcus lactis subsp. lactis and subsp. cremoris. L. monocytogenes strains isolated from four independent food-borne illness incidents were analyzed by all typing methods. Patient and product isolates were not distinguishable by serotyping, ribotyping, or multilocus enzyme electrophoresis. DNA fingerprinting was the only method capable of differentiating these strains, or conversely, of proving relatedness of patient-product pairs of isolates. This method was a relatively simple, sensitive, reproducible, and highly discriminating method for epidemiological tracking of L. monocytogenes implicated in food-borne illness.

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Application of whole-cell DNA restriction endonuclease profiles to the epidemiology of Clostridium difficile-induced diarrhea

Cited by 88 publications

References 19 publications

Phenotypic and Genotypic Diversity of the Flagellin Gene (fliC) among Clostridium difficile Isolates from Different Serogroups

Phenotypic and Genotypic Diversity of the Flagellin Gene (fliC) among Clostridium difficile Isolates from Different Serogroups

Development of a new PCR-ribotyping method for Clostridium difficile based on ribosomal RNA gene sequencing

Comparison of methods for discrimination between strains of Listeria monocytogenes from epidemiological surveys

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