An Update on Clostridium difficile Toxinotyping

Rupnik, Maja; Janežič, Sandra

doi:10.1128/jcm.02083-15

Cited by 101 publications

(81 citation statements)

References 35 publications

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“…Both were already included in the updated toxinotyping scheme [37], but are here described more detailed. Toxinotype IXb (PCR ribotype 244) is similar to the reference strain of toxinotype IX in the main fragments B1 and A3 (most variable regions in tcd B and tcd A, coding for catalytic and binding domain, respectively).…”

Section: Resultsmentioning

confidence: 99%

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Highly Divergent Clostridium difficile Strains Isolated from the Environment

et al. 2016

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Clostridium difficile is one of the most important human and animal pathogens. However, the bacterium is ubiquitous and can be isolated from various sources. Here we report the prevalence and characterization of C. difficile in less studied environmental samples, puddle water (n = 104) and soil (n = 79). C. difficile was detected in 14.4% of puddle water and in 36.7% of soil samples. Environmental strains displayed antimicrobial resistance patterns comparable to already published data of human and animal isolates. A total of 480 isolates were grouped into 34 different PCR ribotypes. More than half of these (52.9%; 18 of 34) were already described in humans or animals. However, 14 PCR ribotypes were new in our PCR ribotype library and all but one were non-toxigenic. The multilocus sequence analysis of these new PCR ribotypes revealed that non-toxigenic environmental isolates are phylogenetically distinct and belong to three highly divergent clades, two of which have not been described before. Our data suggest that environment is a potential reservoir of genetically diverse population of C. difficile.

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Section: Resultsmentioning

confidence: 99%

“…Another new toxinotype, XId (PCR ribotype 153(CE)), differed from other toxinotype XI strains (XIa-c) in different RFLP pattern of A3 fragment of tcdA gene. A truncated PaLoc, with just a part of 3’ end present (equivalent to A3 fragment), characterizes toxinotype XI strains, corresponding to A-B-CDT+ phenotype [37]. …”

Section: Resultsmentioning

confidence: 99%

Highly Divergent Clostridium difficile Strains Isolated from the Environment

et al. 2016

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“…Due to the large size of the toxins, and the difficulty in resolving the repeats in tcdA, the study of such variation has relied mostly on the PCR-RFLP-based method toxinotyping, which currently describes 34 toxinotypes (I-XXXIV) (Rupnik and Janezic, 2016). Variations with the most visible effects are , 2015).…”

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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“…Full-length wild-type TcdA and TcdB from VPI 10463 strain (RT 087, toxinotype 0 [23]), and TcdB from strain M68 (TcdBM68, RT 017, toxinotype VIII [23]) were recombinant toxins expressed in Bacillus megaterium and purified from total crude extracts using the same method as described previously [24]. TcdB from a clinical isolate [25] (TcdB027, RT 027, toxinotype III [23]) was a gift from Dr. Xinhua Chen at Harvard University.…”

Section: Methodsmentioning

confidence: 99%

“…TcdB from a clinical isolate [25] (TcdB027, RT 027, toxinotype III [23]) was a gift from Dr. Xinhua Chen at Harvard University.…”

Section: Methodsmentioning

confidence: 99%

The role of purified Clostridium difficile glucosylating toxins in disease pathogenesis utilizing a murine cecum injection model

et al. 2017

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Most pathogenic Clostridium difficile produce two major exotoxins TcdA and TcdB, in the absence of which the bacterium is non-pathogenic. While it is important to investigate the role of each toxin in the pathogenesis of C. difficile infection (CDI) using isogenic strains, it is impossible to precisely control the expression levels of individual toxins and exclude bacterial factors that may contribute to the toxins' effects during infection. In this study, we utilized an acute intestinal disease model by injecting purified toxins directly into mouse cecum after a midline laparotomy. We evaluated the physical condition of mice by clinical score and survival, and the intestinal tissue damage and inflammation by histology. Depending on the dose of the toxins, mice developed mild to severe colitis, experienced diarrhea or rapidly died. We found that both purified TcdA and TcdB were able to induce clinical disease, intestinal inflammation, and tissue damage that resembled CDI. TcdA was significantly faster in inducing intestinal inflammation and tissue damage, and was approximately five times more potent than TcdB in terms of inducing severe gut disease and death outcomes in mice. Moreover, we found that the two toxins had significant synergistic effects on disease induction. Comparison of the in vivo toxicity of TcdB from clinical strains revealed that TcdB from an epidemic RT 027 strain was more toxic than the others. Our study thus demonstrates that both TcdA and TcdB, independent of other factors from C. difficile bacterium, are able to cause disease that resembles CDI and highlights the importance of targeting both toxins for vaccines and therapeutics against the disease.

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An Update on Clostridium difficile Toxinotyping

Cited by 101 publications

References 35 publications

Highly Divergent Clostridium difficile Strains Isolated from the Environment

Highly Divergent Clostridium difficile Strains Isolated from the Environment

Clostridium difficile infection: Evolution, phylogeny and molecular epidemiology

The role of purified Clostridium difficile glucosylating toxins in disease pathogenesis utilizing a murine cecum injection model

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