Molecular Analysis of the Pathogenicity Locus and Polymorphism in the Putative Negative Regulator of Toxin Production (TcdC) among
            <i>Clostridium difficile</i>
            Clinical Isolates

Spigaglia, Patrizia; Mastrantonio, Paola

doi:10.1128/jcm.40.9.3470-3475.2002

Cited by 258 publications

(229 citation statements)

References 35 publications

(40 reference statements)

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“…PCR products were separated by electrophoresis on a 1.5 % agarose gel and visualized with ethidium bromide staining and the images captured using Alpha Imager software (Alpha Innotech). The presence of deletions or mutations in the tcdC gene was investigated by PCR amplification of the tcdC gene by following the methods outlined by Spigaglia & Mastrantonio (2002). PCR products were purified and sequenced.…”

Section: Methodsmentioning

confidence: 99%

Multidrug-resistant North American pulsotype 2 Clostridium difficile was the predominant toxigenic hospital-acquired strain in the province of Manitoba, Canada, in 2006–2007

Karlowsky

Zhanel

Hammond

et al. 2012

Journal of Medical Microbiology

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The objective of the current study was to determine whether the antimicrobial susceptibility profile or genotype of hospital-acquired isolates of Clostridium difficile differed from isolates causing community-acquired disease. Five hundred diarrhoeal stool samples (one .2 ml sample per patient) from patients across Manitoba, Canada, in 2006-2007 that were reported as C. difficile toxin positive were cultured, resulting in 432 isolates of toxin-positive C. difficile for analysis. Of these 432 isolates, acquisition status could be determined for 235 (54.4 %); 182 (77.4 %) isolates were hospital acquired and 53 (22.6 %) were community acquired. North American pulsotype (NAP) designations based on SmaI PFGE could be defined for 52.3 % of the 432 isolates, with NAP2 (n5122) being the most common. Ninety-one per cent (71/78) of NAP2 isolates were recovered from patients with hospital-acquired C. difficile disease. Other NAP types and isolates with non-NAP-type PFGE patterns were less frequently associated with hospitalacquired disease. Community-acquired disease (35.3 % of isolates) was associated with a wide variety of NAP types. NAP2 isolates were homogeneous (85.5 % had SmaI PFGE pattern 0003) and demonstrated low susceptibility to moxifloxacin (6.6 %) and clindamycin (1.6 %) compared with non-NAP2 isolates (64.1-93.2 % moxifloxacin susceptible; 14.1-28.2 % clindamycin susceptible). All isolates of C. difficile in Manitoba were susceptible to metronidazole, piperacillin-tazobactam, amoxicillin-clavulanate and meropenem. NAP2 isolates of toxigenic C. difficile were approximately three times more common than NAP1 isolates (28.2 vs 9.1 %) in Manitoba in [2006][2007], and these isolates demonstrated high levels of clonality and multidrug resistance, and were associated with hospital acquisition.

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Section: Methodsmentioning

confidence: 99%

Multidrug-resistant North American pulsotype 2 Clostridium difficile was the predominant toxigenic hospital-acquired strain in the province of Manitoba, Canada, in 2006–2007

Karlowsky

Zhanel

Hammond

et al. 2012

Journal of Medical Microbiology

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“…Toxinotyping was performed on a representative of each ribotype (16). Additionally, the tcdC gene from a representative of each ribotype was amplified and sequenced as described previously (18).…”

Section: Methodsmentioning

confidence: 99%

Detection and Enumeration ofClostridium difficileSpores in Retail Beef and Pork

Weese

Avery

Rousseau

et al. 2009

Appl Environ Microbiol

160

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Recent studies have identified Clostridium difficile in food animals and retail meat, and concern has been raised about the potential for food to act as a source of C. difficile infection in humans. Previous studies of retail meat have relied on enrichment culture alone, thereby preventing any assessment of the level of contamination in meat. This study evaluated the prevalence of C. difficile contamination of retail ground beef and ground pork in Canada. Ground beef and ground pork were purchased from retail outlets in four Canadian provinces. Quantitative and enrichment culture was performed. Clostridium difficile was isolated from 28/230 (12%) samples overall: 14/115 (12%) ground beef samples and 14/115 (12%) ground pork samples (P ‫؍‬ 1.0). For ground beef, 10/14 samples (71%) were positive by enrichment culture only. Of the 4 ground beef samples that were positive by direct culture, 20 spores/g were present in 2 while 120 and 240 spores/g were present in 1 each. For ground pork, 10/14 (71%) samples were positive by enrichment culture only. Of the 4 ground pork samples that were positive by direct culture, 20 spores/g were present in 3 while 60 spores/g were present in 1. Ribotype 078 predominated, consistent with some previous studies of C. difficile in food animals. Ribotype 027/North American pulsotype 1 was also identified in both retail beef and pork. This study has identified relatively common contamination of retail ground beef and pork with C. difficile spores; however, the levels of contamination were very low.

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“…Since 2003, outbreaks of C. difficile infection (CDI) due to an emerging strain of PCR ribotype 027 possessing CDT and resistant to erythromycin and/or clindamycin and newer fluoroquinolones have been reported in North America and Europe (Loo et al, 2005;Kuijper et al, 2008;Clements et al, 2010). This strain has a point mutation in tcdC, a putative negative regulator of toxins A and B (Spigaglia & Mastrantonio, 2002;McDonald et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Characterization and antimicrobial susceptibility of Clostridium difficile strains isolated from adult patients with diarrhoea hospitalized in two university hospitals in Poland, 2004–2006

Pituch

Obuch-Woszczatyński

Wultańska

et al. 2011

Journal of Medical Microbiology

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Characterization and antimicrobial susceptibility of Clostridium difficile strains isolated from adult patients with diarrhoea hospitalized in two university hospitals in Poland, 2004Poland, -2006 This study analysed 330 Clostridium difficile strains isolated from patients with C. difficile infection who were hospitalized in two university hospitals (H1 and H2) in Warsaw, Poland, over the period [2004][2005][2006]. Strains were investigated for the presence of tcdA (A), tcdB (B) and binary toxin (CDT) genes, and antimicrobial susceptibility was determined against nine agents. Among the 330 C. difficile isolates, 150 (45.4 %) were classified as A + B + CDT " , 18 (5.5 %) as A + B + CDT + , 144 (43.6 %) as A " B + CDT " and 18 (5.5 %) as A " B " CDT " . The predominant PCR ribotype in hospitals H1 and H2 was type 017 and accounted for 48.3 and 40.0 %, respectively. Only one PCR ribotype 027 strain was found. The rates of resistance to erythromycin and clindamycin in hospitals H1 and H2 were 53.6 and 53.6 %, and 48.6 and 47.5 %, respectively, whereas resistance rates to the newer fluoroquinolones gatifloxacin and moxifloxacin were 38.5 and 38.5 % (H1) and 38.4 and 40.1 % (H2). Erythromycin resistance was frequently associated with resistance to clindamycin and newer fluoroquinolones in strains belonging to type 017. No metronidazole-and vancomycin-resistant isolates were found, although two C. difficile isolates had elevated MIC values of metronidazole (MIC range 1.0"1.5 mg l "1 ) and 15 strains revealed elevated MIC values for vancomycin (MIC range 1.5-2.0 mg l "1 ). In conclusion, an increase in non-027 CDT-producing C. difficile strains was observed in Poland, but C. difficile PCR ribotype 017 remains a major circulating type. INTRODUCTIONClostridium difficile is a major cause of antibioticassociated diarrhoea in hospitalized patients (Freeman et al., 2010). Toxigenic isolates usually produce two toxins: toxin A (A) and toxin B (B). A 2 B + strains do not produce detectable amounts of toxin A due to a deletion in the repeating sequence of the tcdA gene encoding the toxin (Kato et al., 1998). Some C. difficile strains produce a third additional toxin, called C. difficile binary toxin (CDT), which can enhance the attachment of C. difficile to intestinal epithelial cells (Schwan et al., 2009)

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Molecular Analysis of the Pathogenicity Locus and Polymorphism in the Putative Negative Regulator of Toxin Production (TcdC) among Clostridium difficile Clinical Isolates

Cited by 258 publications

References 35 publications

Multidrug-resistant North American pulsotype 2 Clostridium difficile was the predominant toxigenic hospital-acquired strain in the province of Manitoba, Canada, in 2006–2007

Multidrug-resistant North American pulsotype 2 Clostridium difficile was the predominant toxigenic hospital-acquired strain in the province of Manitoba, Canada, in 2006–2007

Detection and Enumeration ofClostridium difficileSpores in Retail Beef and Pork

Characterization and antimicrobial susceptibility of Clostridium difficile strains isolated from adult patients with diarrhoea hospitalized in two university hospitals in Poland, 2004–2006

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