Detection of Clostridium difficile toxin in various tissue culture monolayers

Maniar, A. C.; Williams, T.; Hammond, Gregory

doi:10.1128/jcm.25.10.1999-2000.1987

Cited by 32 publications

(10 citation statements)

References 3 publications

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“…The cytotoxicity test and toxigenic culture remained the most sensitive (>92%) and specific (>99%) methods. Nevertheless, the results of the cytotoxicity assay depend, to some extent, on the cell line, the age of the cells, the way in which stool specimens are processed, and the test format; moreover, they can sometimes be limited by interfering substances in the stool specimens (21,28).…”

Section: Discussionmentioning

confidence: 99%

Comparison of three enzyme immunoassays, a cytotoxicity assay, and toxigenic culture for diagnosis of Clostridium difficile-associated diarrhea

et al. 1993

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Enzyme immunoassays (EIAs) based on monoclonal antibodies for the detection of Clostridium difficile toxins have recently been developed for clinical use. The aim of this study was to compare three commercially available EIAs, two for toxin A (Premier C. difficile Toxin A; Meridian, Osi, Elancourt, France; and Vidas C. difficik Toxin A; bioMerieux, Marcy l'Etoile, France) and one for toxins A and B (Cytoclone A + B EIA; Cambridge Biotech Corp., Codiapharm, Evian, France), with a cytotoxicity assay and toxigenic culture for the diagnosis of C. difficie-associated diarrhea (CDAD). The study was performed with 285 fresh stools from 285 patients with suspected CDAD. In case of disagreement, the tests were repeated on a frozen aliquot of the same stool sample, and the patient's chart was reviewed. CDAD diagnosis was established in 55 cases (incidence, 19.3%). The sensitivities and specificities of the methods were, respectively, 92.7 and 100% for the cytotoxicity assay, 96.4 and 99.1% for toxigenic culture, 75.5 and 97.8% for Cytoclone, 65.4 and 99.6% for Premier, and 65.4 and 100% for Vidas. The results were uninterpretable in 3.2% of cases with Cytoclone, 0.3% with Premier, and 2.5% with Vidas. We conclude that the cytotoxicity assay and toxigenic culture remain the best methods for the diagnosis of CDAD even though they lack standardization and require 48 to 96 h to obtain the result. Despite their rapidity and simplicity, EIAs are not sensitive enough to be relied on as the sole laboratory test.

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Section: Discussionmentioning

confidence: 99%

Comparison of three enzyme immunoassays, a cytotoxicity assay, and toxigenic culture for diagnosis of Clostridium difficile-associated diarrhea

et al. 1993

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“…False-negative tests may reflect undetectable quantities of cytotoxin in patients with early and mild disease, the production of toxins not identified by the assay, inactivation of toxin during transport, 32 and decreased toxin sensitivity of certain cell lines. 33 Positive tests in the absence of disease may occur in up to 5% of healthy adults and hospital control patients. 25,34 The predictive models in this study showed limited ability to discriminate positive from negative outcomes (as indicated by ROC curve areas in the 0.6-0.7 range).…”

Section: Discussionmentioning

confidence: 99%

Predicting Clostridium difficile Stool Cytotoxin Results in Hospitalized Patients with Diarrhea

et al. 1997

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OBJECTIVE: To validate a model for the prediction of Clostridium difficile cytotoxin assay results, and to identify a subgroup of patients with a very low likelihood of C. difficile -associated disease in whom the yield of routine cytotoxin testing is low. DESIGN:Prospective cohort study. Relevant clinical symptoms, signs, and antibiotic exposure were recorded before reporting of assay results. Each predictor was assigned a score based on regression coefficients, and patients were stratified according to their total score. SETTING:Two urban, tertiary care, university hospitals. PATIENTS:A total of 609 consecutive adult inpatients who received testing for C. difficile cytotoxin during a 3-month period in 1994. MEASUREMENTS AND MAIN RESULTS:The prevalence of positive cytotoxin assays was 8% in the validation set, compared with 14% in the derivation set. Defining patients without both prior antibiotic use and at least one symptom predictor (significant diarrhea or abdominal pain) as a low-risk subgroup, the misclassification rate was 2.8% (5/177) for assay results; of the five misclassified cases patients, only one was judged to have C. difficile -associated disease. Use of this rule to identify low-risk patients could have potentially averted 29% of all cytotoxin assays. lostridium difficile accounts for substantial morbidity in hospitalized patients and has been recognized to be the most common cause of antibiotic-associated colitis. 1 Disease associated with C. difficile is also costly. In one study, C. difficile colitis during hospitalization accounted for $2,000 in additional charges per patient. 2 Testing for C. difficile accounted for nearly 68% of total hospital charges associated with the laboratory evaluation of diarrhea in another study. 3 The most widely used method for diagnosis of C. difficile -associated disease is based on detecting a cytotoxin produced by C. difficile by means of a tissue culture assay. 4,5 The yield of C. difficile toxin testing in hospitalized patients with diarrhea has been reported to be approximately 20% in patients hospitalized longer than 3 days. [6][7][8][9] Other investigators, however, have suggested that approximately 40% of testing for C. difficile could be eliminated by using selective criteria. 10 Moreover, the yield of serial (repeated) tests for C. difficile toxin has been reported to be low. 11 In a retrospective study of consecutive patients who underwent first-time testing for C. difficile cytotoxin at a university medical center, where the overall prevalence of positive cytotoxin assays was 14%, Katz et al. developed a simple three-variable decision rule. 5 This rule identified a clinical subgroup at low risk of positive cytotoxin assay results (patients without the combination of antibiotic use within 30 days prior to testing and either a significant diarrhea or abdominal pain). This subgroup accounted for 37% of all cytotoxin assays performed. CONCLUSIONS:Although the subjects of this earlier study had a broad range of symptoms and exposure histories, they wer...

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“…Furthermore, our sensitivity of 80% was comparable to previously reported rates . Many factors including final dilution of inoculum, age and type of the cell line employed influence CTA results (23)(24)(25). As we batched our specimens and the cell cultures were fresh , it is doubtful that the res ults of CTA could be duplicated in the routine laboratmy setting.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Culture, Cytotoxin Assay and Two Eia Tests with Clinical Diagnosis of Clostridium difficile‐Associated Diarrhea

Binning

John

Schieven

et al. 1993

Canadian Journal of Infectious Diseases and Medical Microbiolog

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Detection of Clostridium difficile toxin in various tissue culture monolayers

Cited by 32 publications

References 3 publications

Comparison of three enzyme immunoassays, a cytotoxicity assay, and toxigenic culture for diagnosis of Clostridium difficile-associated diarrhea

Comparison of three enzyme immunoassays, a cytotoxicity assay, and toxigenic culture for diagnosis of Clostridium difficile-associated diarrhea

Predicting Clostridium difficile Stool Cytotoxin Results in Hospitalized Patients with Diarrhea

Comparison of Culture, Cytotoxin Assay and Two Eia Tests with Clinical Diagnosis of Clostridium difficile‐Associated Diarrhea

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