Detection of virulence genes of <i>Clostridium difficile</i> by multiplex PCR

Antikainen, Jenni; Pasanen, Tanja; Mero, Sointu; Tarkka, Eveliina; Kirveskari, Juha; Kotila, Saara; Mentula, Silja; Könönen, Eija; Virolainen-Julkunen, Anni; Vaara, Martti; Tissari, Päivi

doi:10.1111/j.1600-0463.2009.02509.x

Cited by 34 publications

(19 citation statements)

References 23 publications

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“…This may well explain why several studies have reported that an aberrant tcdC genotype does not predict increased toxin production or, indeed, increased virulence (4,6,26,32,39). Notwithstanding this, it should be emphasized that the ⌬117 tcdC frameshift mutation still provides a convenient basis upon which to make a presumptive identification of PCR ribotype 027 strains (1,3,9,40), even though this itself does not provide a broadly applicable rationale for the perceived notion that PCR ribotype 027 strains are "high-level" toxin producers.…”

Section: Discussionmentioning

confidence: 99%

Precise Manipulation of the Clostridium difficile Chromosome Reveals a Lack of Association between the tcdC Genotype and Toxin Production

Cartman

Kelly

Heeg

et al. 2012

Appl Environ Microbiol

221

226

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Clostridium difficile causes a potentially fatal diarrheal disease through the production of its principal virulence factors, toxin A and toxin B. The tcdC gene is thought to encode a negative regulator of toxin production. Therefore, increased toxin production, and hence increased virulence, is often inferred in strains with an aberrant tcdC genotype. This report describes the first allele exchange system for precise genetic manipulation of C. difficile, using the codA gene of Escherichia coli as a heterologous counterselection marker. It was used to systematically restore the ⌬117 frameshift mutation and the 18-nucleotide deletion that occur naturally in the tcdC gene of C. difficile R20291 (PCR ribotype 027). In addition, the naturally intact tcdC gene of C. difficile 630 (PCR ribotype 012) was deleted and then subsequently restored with a silent nucleotide substitution, or "watermark," so the resulting strain was distinguishable from the wild type. Intriguingly, there was no association between the tcdC genotype and toxin production in either C. difficile R20291 or C. difficile 630. Therefore, an aberrant tcdC genotype does not provide a broadly applicable rationale for the perceived notion that PCR ribotype 027 strains are "high-level" toxin producers. This may well explain why several studies have reported that an aberrant tcdC gene does not predict increased toxin production or, indeed, increased virulence.

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Section: Discussionmentioning

confidence: 99%

Precise Manipulation of the Clostridium difficile Chromosome Reveals a Lack of Association between the tcdC Genotype and Toxin Production

Cartman

Kelly

Heeg

et al. 2012

Appl Environ Microbiol

221

226

View full text Add to dashboard Cite

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“…Lem ee et al (2004) [140] designed a multiplex PCR for the simultaneous identification and toxigenic type characterisation of C. difficile isolates. Several other studies have proposed different multiplex PCR primers and protocols not only to detect the genes encoding the major toxins A and B but also to detect binary toxin genes (cdtA and cdtB) and other deletions in the Paloc genes [174].…”

Section: Difficile Typing Methodsmentioning

confidence: 99%

Clostridium difficile infection: Early history, diagnosis and molecular strain typing methods

Rodríguez

Broeck

Taminiau

et al. 2016

Microbial Pathogenesis

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a b s t r a c tRecognised as the leading cause of nosocomial antibiotic-associated diarrhoea, the incidence of Clostridium difficile infection (CDI) remains high despite efforts to improve prevention and reduce the spread of the bacterium in healthcare settings. In the last decade, many studies have focused on the epidemiology and rapid diagnosis of CDI. In addition, different typing methods have been developed for epidemiological studies. This review explores the history of C. difficile and the current scope of the infection. The variety of available laboratory tests for CDI diagnosis and strain typing methods are also examined.

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“…All unique strains were compared with the Cardiff-European Center for Disease Control (ECDC) collection of C. difficile strains; a Washington University (WU) strain number was assigned to unique strains that did not match any in the Cardiff-ECDC collection. All C. difficile isolates were also tested by PCR for the presence of tcdA and tcdB and binary toxin genes by PCR, as previously described (23,31).…”

Section: Methodsmentioning

confidence: 99%

Risk Factors for Acquisition and Loss of Clostridium difficile Colonization in Hospitalized Patients

Dubberke

Reske

Seiler

et al. 2015

Antimicrob Agents Chemother

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bAsymptomatic colonization may contribute to Clostridium difficile transmission. Few data identify which patients are at risk for colonization. We performed a prospective cohort study of C. difficile colonization and risk factors for C. difficile acquisition and loss in hospitalized patients. Patients admitted to medical or surgical wards at a tertiary care hospital were enrolled; interviews and chart review were performed to determine patient demographics, C. difficile infection (CDI) history, medications, and health care exposures. Stool samples/rectal swabs were collected at enrollment and discharge; stool samples from clinical laboratory tests were also included. Samples were cultured for C. difficile, and the isolates were tested for toxins A and B and ribotyped. Chi-square tests and univariate logistic regression were used for the analyses. Two hundred thirty-five patients were enrolled. Of the patients, 21% were colonized with C. difficile (toxigenic and nontoxigenic) at admission and 24% at discharge. Ribotype 027 accounted for 6% of the strains at admission and 12% at discharge. Of the patients colonized at admission, 78% were also colonized at discharge. Cephalosporin use was associated with C. difficile acquisition (47% of patients who acquired C. difficile versus 25% of patients who did not; P ‫؍‬ 0.03). ␤-lactam-␤-lactamase inhibitor combinations were associated with a loss of C. difficile colonization (36% of patients who lost C. difficile colonization versus 8% of patients colonized at both admission and discharge; P ‫؍‬ 0.04), as was metronidazole (27% versus 3%; P ‫؍‬ 0.03). Antibiotic use affects the epidemiology of asymptomatic C. difficile colonization, including acquisition and loss, and it requires additional study.C lostridium difficile infection (CDI) is the most common infectious cause of hospital-associated diarrhea, and while most CDI infections are mild, severe CDI can lead to outcomes, such as toxic megacolon, colectomy, and death (1-4). Current estimates of the excess health care costs associated with CDI in the United States are $4.8 billion per year (5). The emergence of the epidemic NAP1/BI/027 strain of C. difficile (6-8) and increased infection rates (9, 10) have generated a renewed interest in this pathogen.Despite this interest in CDI, optimal methods for preventing CDI remain poorly understood. Traditionally, symptomatic CDI patients have been considered the primary reservoir for C. difficile transmission because they shed more C. difficile in their stool than asymptomatically colonized patients (11-13). However, recently published studies indicate only one-third or fewer of new CDI cases in the hospital setting can be attributed to transmission from another CDI case (14-17). It is possible that asymptomatic carriers are an important source of C. difficile transmission (18, 19). Curry et al. (20) found that the percentage of CDI cases related to symptomatic CDI patients and that related to asymptomatic C. difficile carriers was almost identical (30% versus 29%, respectively...

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Detection of virulence genes of Clostridium difficile by multiplex PCR

Cited by 34 publications

References 23 publications

Precise Manipulation of the Clostridium difficile Chromosome Reveals a Lack of Association between the tcdC Genotype and Toxin Production

Precise Manipulation of the Clostridium difficile Chromosome Reveals a Lack of Association between the tcdC Genotype and Toxin Production

Clostridium difficile infection: Early history, diagnosis and molecular strain typing methods

Risk Factors for Acquisition and Loss of Clostridium difficile Colonization in Hospitalized Patients

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