Use of an enzyme-linked immunoassay for Clostridium difficile serogrouping

Delmée, Michel; Depitre, C.; Corthier, Gérard; Ahoyo, Angèle; Avesani, Véronique

doi:10.1128/jcm.31.9.2526-2528.1993

Cited by 18 publications

(5 citation statements)

References 13 publications

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“…Ninety-nine strains of C. difficile were used for this study. They included 20 reference strains kindly supplied by M. Delmée (University of Louvain, Brussels, Belgium) and corresponding to the 20 different serogroups (9) (11), and 4 pairs of epidemiologically related strains. Strains were cultured on TCCA plates (brain heart infusion agar supplemented with 5% defibrinated horse blood, 0.1% taurocholate, cefoxitin at 10 g ml Ϫ1 , and cycloserine at 250 g ml Ϫ1 ) and incubated in an anaerobic atmosphere.…”

Section: Methodsmentioning

confidence: 99%

“…Colonies were identified by use of an enzymatic profile from a RapID 32 A gallery (bioMérieux, La Balme les Grottes, France). Serogrouping was performed by use of an enzyme-linked immunosorbent assay according to the method described by Delmée et al (9) with antisera A1, A5, A8, A9, A10, C, D, F, G, H, and K. The distribution of sporadic and related strains among the different serogroups is shown in Table 1.…”

Section: Methodsmentioning

confidence: 99%

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Comparison of PCR-Ribotyping, Arbitrarily Primed PCR, and Pulsed-Field Gel Electrophoresis for Typing Clostridium difficile

et al. 2000

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Clostridium difficile is now recognized as the major agent responsible for nosocomial diarrhea in adults. Among the genotyping methods available, arbitrarily primed PCR (AP-PCR), PCR-ribotyping, and pulsed-field gel electrophoresis (PFGE) have been widely used for investigating outbreaks of C. difficileinfections. However, the comparative typing ability, reproducibility, discriminatory power, and efficiency of these methods have not been fully investigated. We compared the results of three methods—AP-PCR with three different primers (AP3, AP4, and AP5), PCR-ribotyping, and PFGE (with SmaI endonuclease)—to differentiate 99 strains of C. difficile that had been previously serogrouped. Typing abilities were 100% for PCR-ribotyping and AP-PCR with AP3 and 90% for PFGE, due to early DNA degradation in strains from serogroup G. Reproducibilities were 100% for PCR-ribotyping and PFGE but only 88% for AP-PCR with AP3, 67% for AP-PCR with AP4, and 33% for AP-PCR with AP5. Discriminatory power for unrelated strains was >0.95 for all the methods but was lower for PCR-ribotyping among serogroups D and C. PCR-based methods were easier and quicker to perform, but their fingerprints were more difficult to interpret than those of PFGE. We conclude that PCR-ribotyping offers the best combination of advantages as an initial typing tool for C. difficile.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Comparison of PCR-Ribotyping, Arbitrarily Primed PCR, and Pulsed-Field Gel Electrophoresis for Typing Clostridium difficile

et al. 2000

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“…Serotyping. Serotyping of the C. difficile strains was performed by the method of Delmée et al (14). Eleven antisera specific for serogroups A1, A5, A8, A9, A10, C, D, F, G, H, and K were used in an enzyme-linked immunosorbent assay format, as described previously (14).…”

Section: Methodsmentioning

confidence: 99%

Antimicrobial Susceptibilities and Serogroups of Clinical Strains of Clostridium difficile Isolated in France in 1991 and 1997

Barbut

Decré

Burghoffer

et al. 1999

Antimicrob Agents Chemother

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Glycopeptides (vancomycin and teicoplanin) and metronidazole are the drugs of choice for the treatment of Clostridium difficile infections, but trends in susceptibility patterns have not been assessed in the past few years. The objective was to study the MICs of glycopeptides and metronidazole for unrelated C. difficile strains isolated in 1991 (n = 100) and in 1997 (n = 98) by the agar macrodilution, the E-test, and the disk diffusion methods. Strain susceptibilities to erythromycin, clindamycin, tetracycline, rifampin, and chloramphenicol were also determined by the ATB ANA gallery (bioMérieux, La Balme-les-Grottes, France). The MICs at which 50% of isolates are inhibited (MIC50s) and MIC90s of glycopeptides and metronidazole remained stable between 1991 and 1997. All the strains were inhibited by concentrations that did not exceed 2 μg/ml for vancomycin and 1 μg/ml for teicoplanin. Comparison of MICs determined by the agar dilution method recommended by the National Committee for Clinical Laboratory Standards and the E test showed correlations (±2 dilutions) of 86.6, 95.9, and 99% for metronidazole, vancomycin, and teicoplanin, respectively. The E test always underestimated the MICs. Strains with decreased susceptibility to metronidazole (MICs, ≥8 μg/ml) were isolated from six patients (n = 4 in 1991 and n = 2 in 1997). These strains were also detected by the disk diffusion method (zone inhibition diameter, ≤21 mm); they belonged to nontoxigenic serogroup D (n = 5) and toxigenic serogroup H (n = 1). Decreased susceptibility to erythromycin (MICs, ≥1 μg/ml), clindamycin (MICs, ≥2 μg/ml), tetracycline (MICs, ≥8 μg/ml), rifampin (MICs, ≥4 μg/ml), and chloramphenicol (MICs, ≥16 μg/ml) was observed in 64.2, 80.3, 23.7, 22.7, and 14.6% of strains, respectively. Strains isolated in 1997 were more susceptible than those isolated in 1991, and this trend was correlated to a major change in serogroup distribution. Periodic studies are needed in order to detect changes in serogroups and the emergence of strains with decreased susceptibility to therapeutic drugs.

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“…Serogrouping of C. difficile strains was performed according to the method of Delmee et al 2 ' Eleven antisera specific for serogroups Al, A5, A8, A9, A10, C, D, F, G, H, and K were used in an enzyme-linked immunosorbent assay, as described elsewhere. 21 Genetic analysis. DNA was extracted with the Instagene Matrix kit (Bio-Rad).…”

Section: Definitionsmentioning

confidence: 99%

Clinical Features ofClostridium difficile–Associated Infections and Molecular Characterization of Strains: Results of a Retrospective Study, 2000-2004

Barbut

Gariazzo

Bonné

et al. 2007

Infect. Control Hosp. Epidemiol.

151

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Use of an enzyme-linked immunoassay for Clostridium difficile serogrouping

Cited by 18 publications

References 13 publications

Comparison of PCR-Ribotyping, Arbitrarily Primed PCR, and Pulsed-Field Gel Electrophoresis for Typing Clostridium difficile

Comparison of PCR-Ribotyping, Arbitrarily Primed PCR, and Pulsed-Field Gel Electrophoresis for Typing Clostridium difficile

Antimicrobial Susceptibilities and Serogroups of Clinical Strains of Clostridium difficile Isolated in France in 1991 and 1997

Clinical Features ofClostridium difficile–Associated Infections and Molecular Characterization of Strains: Results of a Retrospective Study, 2000-2004

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