A Novel Toxinotyping Scheme and Correlation of Toxinotypes with Serogroups of
            <i>Clostridium difficile</i>
            Isolates

Rupnik, Maja; Avesani, Véronique; Janc, M.; Eichel‐Streiber, Christoph von; Delmée, Michel

doi:10.1128/jcm.36.8.2240-2247.1998

Cited by 298 publications

(138 citation statements)

References 31 publications

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“…This observation indicates that some intrinsic di¡erences in the structural composition of the peptidoglycan layer may exist between the two strains of C. di¤cile. A similar observation in the form of polymorphisms has been found to exist in both tcdA and tcdB genes among the various pathogenic strains of C. di¤cile [18].…”

Section: Genomic Dna Extractionsupporting

confidence: 77%

Sequence analysis of a new open reading frame located in the pathogenicity locus of Clostridium difficile strain 8864

Song¹

1999

FEMS Microbiology Letters

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Strain 8864 is a natural isolate of Clostridium difficile that is toxin B-positive and toxin A-negative. Recent work showed that there is a genetic rearrangement occurring at the pathogenicity locus (PaLoc) of the bacteria. Our investigation in the PaLoc region revealed an open reading frame (tcdF) of 543 bp DNA not reported before. This tcdF could encode a putative polypeptide of 22 kDa. Although no peptide homology was found with other known proteins, we postulate that it could be a novel protein because not only highly consensus ribosome-binding and promoter sequences were found upstream of tcdF, transcript was also identified at the region occupied by tcdF. ß

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Section: Genomic Dna Extractionsupporting

confidence: 77%

Sequence analysis of a new open reading frame located in the pathogenicity locus of Clostridium difficile strain 8864

Song¹

1999

FEMS Microbiology Letters

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“…C. difficile can be divided into more than 150 ribotypes and 24 toxinotypes [55,67] (http:// www.mf.uni-mb.si/mikro/tox).…”

Section: H a R A C T E R I S T I C S O F C D I F F I C I L E P C mentioning

confidence: 99%

Emergence of Clostridium difficile-associated disease in North America and Europe

Kuijper

Coignard

Tüll

2006

Clinical Microbiology and Infection

752

493

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The clinical spectrum of Clostridium difficile-associated disease (CDAD) ranges from diarrhoea to severe life-threatening pseudomembranous colitis. Although not always associated with previous antibiotic exposure, it is in the majority of cases. CDAD is recognised increasingly in a variety of animal species and in individuals previously not considered to be predisposed. C. difficile can be transmitted via personal contact or environmentally. The role of patients and healthcare workers who are symptom-free but colonised with C. difficile in the intestinal tract is unclear. C. difficile, with more than 150 PCR ribotypes and 24 toxinotypes, has a pathogenicity locus (PaLoc) with genes encoding enterotoxin A (tcdA) and cytotoxin B (tcdB). Genes for the binary toxin are located outside the PaLoc, but the role of this toxin is unclear. The recently completed genome sequence of C. difficile 630 revealed a large proportion of 11% of mobile genetic elements, mainly in the form of conjugative transposons. Diagnostic assays include tests for the detection of C. difficile products or genes and culture methods for isolation of a toxin-producing bacterium. Enzyme immunoassays to detect toxin in faeces are widely available, with varying sensitivities and specificities. Despite practical drawbacks and sensitivity less than 100%, the cell cytototoxicity assay is still considered to be the standard. Rapid diagnostic assays are available on a limited scale and require much improvement. Molecular tests enable the detection of carriers of toxigenic and non-toxigenic strains, as does culture. It is highly recommended to culture C. difficile from toxin-positive faeces samples and to store isolates for future characterisation and typing. The financial impact of CDAD on the healthcare system is substantial (5-15,000 euro/case in England and $1.1 billion/year in the USA). Assuming a European Union population of 457 million, the potential cost of CDAD can be estimated to be 3000 million euro/year, and is expected to almost double over the next four decades. In North America, increasing rates of CDAD have been reported in Canada and the USA since March 2003, involving a more severe course, higher mortality, increased risk of relapse and more complications. This increased virulence is presumably associated with higher levels of toxin production by fluoroquinolone-resistant strains belonging to PCR ribotype 027, pulsed-field gel electrophoresis (PFGE) type NAP1, REA (restriction endonuclease analysis) type BI and toxinotype III. In Europe, outbreaks of CDAD due to the new, highly virulent strain of C. difficile PCR ribotype 027, toxinotype III have been recognised in 75 hospitals in England, 16 hospitals in The Netherlands, 13 healthcare facilities in Belgium and nine healthcare facilities in France. These outbreaks are very difficult to control, and preliminary results from case-control studies indicate a correlation with fluoroquinolones and cephalosporins. Information concerning community-acquired cases of ribotype 027 is lacking, and data...

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“…þ a 0, length and restriction patterns of amplified PCR product are the same as in strainVPI10463; R, RFLP detected in amplified fragment; D, deletion detected in amplified fragment; na, PCR fragment can not be amplified. a PCR fragments B1^B3, A1^A3 according to reference [8]. However, to date only a single isolate of toxinotype X is known, although another three similar strains were recently found [16, M. Rupnik and H. Kato, unpublished data] and it seems that this type of variant strain is rather rare.…”

Section: How To De T Ec T C Difficile Var I An T S Tr Ain Smentioning

confidence: 99%

How to detect Clostridium difficile variant strains in a routine laboratory

Rupnik

2001

Clinical Microbiology and Infection

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Toxin A-negative, toxin B-positive strains (A-/B+) are the best studied examples of Clostridium difficile variant strains. In addition, there are some other groups of variant C. difficile strains that produce both toxins (A+/B+) or are non-cytotoxic (A-/B-) but differ from the reference strain VPI 10463 in their toxin genes. Here we describe two simple methods (amplification of the tcdA gene and amplification of the binary toxin gene cdtA) which can be used in rapid screening for variant C. difficile strains in collections or in routine laboratories.

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A Novel Toxinotyping Scheme and Correlation of Toxinotypes with Serogroups of Clostridium difficile Isolates

Cited by 298 publications

References 31 publications

Sequence analysis of a new open reading frame located in the pathogenicity locus of Clostridium difficile strain 8864

Sequence analysis of a new open reading frame located in the pathogenicity locus of Clostridium difficile strain 8864

Emergence of Clostridium difficile-associated disease in North America and Europe

How to detect Clostridium difficile variant strains in a routine laboratory

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