Distribution of Clostridium difficile variant toxinotypes and strains with binary toxin genes among clinical isolates in an American hospital

Geric, Barbara; Rupnik, Maja; Gerding, Dale N.; Grabnar, M.; Johnson, Stuart

doi:10.1099/jmm.0.45610-0

Cited by 144 publications

(100 citation statements)

References 39 publications

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“…Among the serogroup A isolates, 16 were toxigenic (toxin A and B positive) and all were toxinotype 0. These results are in accordance with other studies that reported the high frequency of toxinotype 0 among clinical isolates (Geric et al, 2004). No isolate belonged to the hypertoxigenic toxinotype III recently described in North America (Loo et al, 2005).…”

Section: Resultssupporting

confidence: 83%

Molecular characterization of Clostridium difficile clinical isolates in a geriatric hospital

Poîlane

Humeniuk-Ainouz

Durand

et al. 2007

Journal of Medical Microbiology

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The discriminatory potential of a combination of various typing methods was evaluated on a set of 21 Clostridium difficile isolates obtained from symptomatic patients hospitalized in a geriatric unit and 7 non-toxigenic isolates from the same hospital. Isolates were firstly serotyped and toxinotyped. Of the 28 isolates, 19 belonged to serogroup A. PCR-ribotyping and PCR-RFLP on the fliC and slpA genes were then applied to these 19 isolates. The results suggest that the combination of PCR-ribotyping with PCR-RFLP analysis of slpA could be more discriminatory and suitable for studying C. difficile epidemiology.

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Section: Resultssupporting

confidence: 83%

Molecular characterization of Clostridium difficile clinical isolates in a geriatric hospital

Poîlane

Humeniuk-Ainouz

Durand

et al. 2007

Journal of Medical Microbiology

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“…It has been recognized previously that the presence of functional binary toxin genes encoded by the 4.3 kb CDT locus correlates well with variant toxinotypes including A + B + , A 2 B + and A 2 B 2 strains (Stubbs et al, 2000;Rupnik et al, 2001Rupnik et al, , 2003aGeric et al, 2004;Spigaglia & Mastrantonio, 2002;Goncalves et al, 2004). This study presents an overview of all of the known variant toxinotypes and has confirmed that the full-length CDT locus is found in variant toxinotypes with significantly changed LCT toxin genes (shown in bold in Table 1), whereas the truncated version of the CDT locus is predominant in variant toxinotypes with minor changes in the LCT toxin genes (Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…For toxinotypes IV, V, VI, IX, XIa, XIb and XII we tested one additional strain, for toxinotype III two additional strains and for toxinotype VIII seven additional strains. Three strains of toxinotype III were representatives of subtypes IIIa, IIIb and IIIc, which differed in B2s fragment RFLPs (Geric et al, 2004). Two representatives of the so far unpublished toxinotypes determined in this study (XXIII and XXIV) were also included.…”

Section: Methodsmentioning

confidence: 99%

“…A dendrogram of the PaLoc region for the different toxinotypes was constructed based on PCR RFLP patterns in the reference strains of toxinotypes 0-XXIV. For each toxinotype, data on amplification and RFLPs for seven DNA fragments covering the PaLoc region (Braun et al, 1996;Rupnik et al, 1998;Geric et al, 2004) were included in the matrix: B1 cut with HincII and AccI, B2s cut with HindIII and RsaI, B3s cut with HindIII and RsaI, A1 cut with NsiI and PstI, A2 cut with HaeIII, A3 cut with EcoRI and PLC cut with NcoI.…”

Section: Methodsmentioning

confidence: 99%

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Variant forms of the binary toxin CDT locus and tcdC gene in Clostridium difficile strains

Stare

Delmée

Rupnik

2007

Journal of Medical Microbiology

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Variability in the genes for toxin A, toxin B and other pathogenicity locus regions is well known and is the basis for the distribution of Clostridium difficile strains into variant toxinotypes. Previous data have indicated that some C. difficile strains have a non-functional truncated form of the binary toxin (CDT) locus. This study analysed variability in the CDT locus and the presence of deleted tcdC genes in C. difficile strains. A total of 146 strains were screened, including known variant toxinotypes and non-variant A + B + (toxinotype 0) and A " B " C. difficile strains. In all of the strains studied, only two forms of the CDT locus were found: a full-length 4.3 kb fragment encoding the functional binary toxin or a truncated 2.3 kb fragment. Whilst the full-length CDT locus was found almost exclusively in variant toxinotypes, the truncated form was detected in 79 % of toxinotype 0 strains. Non-toxinogenic A " B " strains with a truncated version were not found and only rarely possessed the full-length CDT locus (A " B " CDT + strains). Four different forms of the tcdC gene were found; three represented deleted versions and typically were found in toxinotypes III-VII, XI, XIV-XVI and XXIV. INTRODUCTIONOver the last decade, Clostridium difficile has developed into one of the most important nosocomial pathogens. This Gram-positive, sporulating micro-organism is associated with post-antibiotic intestinal infections, and the prevalence rate and disease severity seem to be rising (McDonald et al., 2006). Additionally, a new highly virulent and multiresistant type (BI/NAP1/027) has emerged recently and is spreading in Canada, the USA, the UK, the Netherlands, Belgium and France (Loo et al., 2005;Pepin et al., 2004;McDonald et al., 2005;Smith, 2005; Joseph et al., 2005;Kuijper et al., 2006;Tachon et al., 2006). In addition to high fluoroquinolone resistance, this epidemic type has some features previously known but uncommon among C. difficile isolates: variant genes for toxins A (TcdA) and B (TcdB), production of the binary toxin (CDT) and deletions in the tcdC gene.Variability in the tcdA and tcdB genes has been well studied and the strains with changed genes have been grouped into toxinotypes (Rupnik et al., 1998). Major changes are defined as deletions and RFLPs in all six PCR fragments covering tcdA and tcdB, whilst minor changes are limited to only one of the PCR fragments (usually the A3 fragment) covering the toxin genes or other pathogenicity locus (PaLoc) regions. To date, 22 toxinotypes have been described and two additional types are reported in this paper. The best-studied and the most widespread are toxinotypes III (including ribotypes 027, 034, 075 and 080) and VIII (including ribotypes 017 and 047) .Binary toxin CDT is detected in some strains in addition to TcdA and TcdB, the primary virulence factors of C. difficile (Popoff et al., 1988;Rupnik et al., 2003a). Binary toxin CDT is composed of two unlinked components, the enzymic (CDTa) and binding (CDTb) components, and acts as an actin-specific AD...

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“…Several intact mobile elements have also been identified within the PaLoc. Toxinotypes XIVa, Xb, XXII and IXc, all belonging to clade 2, possess a large 2000 bp IStron in tcdA (Geric et al, 2004;Mehlig et al, 2001), a type of mobile element first discovered in C. difficile (Hasselmayer et al, 2004b). The clade 3 PaLoc has a stably integrated transposon Tn6218 which does not interfere with toxin production .…”

Section: Other Lct Elementsmentioning

confidence: 99%

Clostridium difficile infection: Evolution, phylogeny and molecular epidemiology

Elliott

Androga

Knight

et al. 2017

Infection, Genetics and Evolution

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Over the recent decades, Clostridium difficile infection (CDI) has emerged as a global public health threat. Despite growing attention, C. difficile remains a poorly understood pathogen, however, the exquisite sensitivity offered by next generation sequencing (NGS) technology has enabled analysis of the genome of C. difficile, giving us access to massive genomic data on factors such as virulence, evolution, and genetic relatedness within C. difficile groups. NGS has also demonstrated excellence in investigations of outbreaks and disease transmission, in both small and large-scale applications. This review summarizes the molecular epidemiology, evolution, and phylogeny of C. difficile, one of the most important pathogens worldwide in the current antibiotic resistance era.

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Distribution of Clostridium difficile variant toxinotypes and strains with binary toxin genes among clinical isolates in an American hospital

Cited by 144 publications

References 39 publications

Molecular characterization of Clostridium difficile clinical isolates in a geriatric hospital

Molecular characterization of Clostridium difficile clinical isolates in a geriatric hospital

Variant forms of the binary toxin CDT locus and tcdC gene in Clostridium difficile strains

Clostridium difficile infection: Evolution, phylogeny and molecular epidemiology

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