Enzyme Immunoassays for Detection of Clostridium difficile Toxins A and B in Fecal Specimens

Laughon, Barbara E.; Viscidi, Raphael P.; Gdovin, Susan L.; Yolken, Robert H.; Bartlett, John G.

doi:10.1093/infdis/149.5.781

Cited by 104 publications

(57 citation statements)

References 24 publications

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“…The difficulty of completely removing cross-reactive antibodies by absorption has been reported by Laughon et al (1984) and Shahrabadi et al (1984) …”

Section: Resultsmentioning

confidence: 95%

“…Five specimens that were negative with ELISA ( (George, 1984;Bartlett, 1981;Bartlett, 1984 & Wilkins, 1982). Enzyme immunoassays using affinity-purified antibodies to toxins A and B have been described (Lyerly, Sullivan & Wilkins, 1983;Laughon et al 1984). The sensitivity of detection of toxin B by ELISA is reported to be less than that ofthe cytotoxicity assay .…”

Section: Resultsmentioning

confidence: 99%

“…Counterimmunoelectrophoresis (CIE) has been the most widely used method (Ryan, Kwasnik & Tilton, 1980;Welch, Menge & Matsen, 1980; Levine, Kennedy & Lamont, 1982;Wu & Fung, 1983;Jarvis et al 1983;Rennie et al 1984;Lyerly, Phelps & Wilkins, 1985). Latex agglutination (Shahrabadi et al 1984;Lyerly, Phelps & Wilkins 1985) and ELISA (Yolken et al 1981; Lyerly, Sullivan & Wilkins 1983;Laughon et al 1984) have not been as widely evaluated. The time of obtaining results from these techniques varies from 5 min to 4 h. ELISA appears to be the most sensitive of these being able to detect very small quantities of toxin Lyerly, Sullivan & Wilkins, 1983 published results indicate a lack of specificity in most laboratories.…”

Section: Introductionmentioning

confidence: 99%

“…Antisera that are commercially available are produced against toxins of C. difficile or against toxins of C. sordellii. All these antisera contain antibodies to many antigens besides antitoxins to toxins A and B of C. difficile (Poxton & Byrne, 1981;West & Wilkins, 1982;Ryan, Kwasnik & Tilton, 1983;Laughon et al 1984;Rennie et al 1984). The specificity of ELISA was enhanced by using absorbed sera as reported here.…”

Section: Introductionmentioning

confidence: 99%

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Detection ofClostridium difficiletoxins by enzyme immunoassay

Krishnan

1986

J. Hyg.

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SUMMARYAn enzyme-linked immunosorbent assay (ELISA) for the rapid diagnosis of antibiotic-associated colitis (AAC) is presented. Commercially available antisera to Clostridium difficile toxins contain antibodies to other antigens found in non-toxigenic C. difficile and other bacteria. Removal ofthese unwanted antibodies by absorption increased the specificity of ELISA for detection of C. difficile toxins.Specimens tested included 40 faecal extracts positive for cytotoxicity from cases of AAC, 30 diarrhoeic and 30 well-formed stools negative for cytotoxicity and 50 culture filtrates of toxigenic and non-toxigenic C. difficile and other clostridial species. Use of absorbed sera reduced false-positive reactions observed with faecal specimens from 23 to 8 %. About 90 % of specimens that were positive by the tissue culture cytotoxicity test were positive by ELISA using the absorbed sera. The relative merits of ELISA and other methods for the rapid diagnosis of AAC are discussed.

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“…The difficulty of completely removing cross-reactive antibodies by absorption has been reported by Laughon et al (1984) and Shahrabadi et al (1984) …”

Section: Resultsmentioning

confidence: 95%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Detection ofClostridium difficiletoxins by enzyme immunoassay

Krishnan

1986

J. Hyg.

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“…In addition, it was shown recently that C. difficile enterotoxin (toxin A) was primarily responsible for the diarrhoea associated with C. difficile infection (Stephen, 1986). A commercially available latex reagent for toxin A does not detect this toxin (Lyerley & Wilkins, 1986) and enzyme-linked immunosorbent assay for toxin A (Laughon et al 1984) is not routinely performed in diagnostic microbiology laboratories. Moreover, the production of both toxins A and B depends on the composition of the growth medium in vitro and, presumably, on the nutritional status of the gut in vivo (Haslam et al 1986).…”

Section: Introductionmentioning

confidence: 99%

Comparison of alcohol shock enrichment and selective enrichment for the isolation ofClostridium difficile

et al. 1987

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SUMMARYTwo enrichment methods were compared for their ability to recover Clostridium difficile from stool samples. One method used selective enrichment in an antibioticcontaining broth followed by detection with a latex particle agglutination (LPA) reagent. The other used enrichment in a non-selective broth following treatment of the specimen with alcohol. With clinical specimens enrichment culture was significantly more successful at detecting C. difficile than direct plating. Alcohol shock enrichment was twice as effective as direct culture, while selective broth enrichment was three times more effective. The use of LPA for screening selective enrichment broths for C. difficile should prove a cost-effective measure as only positive broths (about 20%) require subculture for confirmation.

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Diagnosis and Treatment of Clostridium difficile Colitis

Fekety¹

1993

JAMA

266

122

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Pseudomembranous colitis associated with antibiotic therapy is almost always due to an overgrowth of Clostridium difficile. If untreated, pseudomembranous colitis can lead to severe diarrhea, hypovolemic shock, toxic dilatation of the colon, cecal perforation, hemorrhage, and death. However, C difficile-associated colitis can mimic the more common "benign" antibiotic-associated diarrhea that is not caused by C difficile. An algorithm for diagnosis management of hospitalized patients with antibiotic diarrhea and C difficile colitis is presented in this review. Diagnosis depends on sigmoidoscopy and/or stool tests for C difficile toxins in all patients with antibiotic-associated diarrhea. If the results of these tests are positive, either metronidazole or vancomycin is recommended for treatment of mild illness, and vancomycin is recommended for treatment of severe illness. Oral therapy is always preferred because it is more reliable. In patients with recurrent or relapsing colitis, treatment with either metronidazole or vancomycin is effective for that episode, but novel approaches, such as the oral or rectal introduction of competing nonpathogenic organisms, may prove to be more successful in prevention of relapses.

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Enzyme Immunoassays for Detection of Clostridium difficile Toxins A and B in Fecal Specimens

Cited by 104 publications

References 24 publications

Detection ofClostridium difficiletoxins by enzyme immunoassay

Detection ofClostridium difficiletoxins by enzyme immunoassay

Comparison of alcohol shock enrichment and selective enrichment for the isolation ofClostridium difficile

Diagnosis and Treatment of Clostridium difficile Colitis

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