Clinical Usefulness of Components of the Triage Immunoassay, Enzyme Immunoassay for Toxins A and B, and Cytotoxin B Tissue Culture Assay for the Diagnosis of Clostridium difficile Diarrhea

Massey, Viki; Gregson, Daniel B.; Chagla, Abdul H.; Storey, Miriam; John, Michael; Hussain, Zafar

doi:10.1309/u8at-l52q-60xy-avx6

Cited by 29 publications

(26 citation statements)

References 22 publications

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“…GDH tests, like stool cultures for C. difficile, are unable to differentiate between toxigenic and nontoxigenic strains, and they cannot establish a diagnosis of CDAD, because colonization with C. difficile in hospitalized patients is frequent and CDAD is a toxin-mediated disease. A number of previous studies have shown that testing for GDH is a useful screening test (3,14,16,20), and the present study confirms this observation.…”

supporting

confidence: 90%

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Performance of the TechLab C. DIFF CHEK-60 Enzyme Immunoassay (EIA) in Combination with the C. difficile Tox A/B II EIA Kit, the Triage C. difficile Panel Immunoassay, and a Cytotoxin Assay for Diagnosis of Clostridium difficile -Associated Diarrhea

et al. 2004

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Clostridium difficile-associated diarrhea (CDAD) is the most commonly identified cause of nosocomial diarrhea (10,12,17). The pathogenicity of C. difficile is due to the production of two exotoxins: toxin A, an enterotoxin, and toxin B. Both toxins A and B contribute to human disease (19).TechLab Inc. (Blacksburg, Va.) has recently marketed an enzyme immunoassay, C. DIFF CHEK-60, which detects glutamate dehydrogenase (GDH), a common C. difficile antigen (the assay is hereinafter referred to as TL-GDH), and also markets an enzyme immunoassay, C. difficile Tox A/B II (Tox-A/B), for the detection of toxins A and B. The aim of this study was to evaluate and compare these two assays with another immunoassay, the Triage C. difficile Panel assay (Biosite Diagnostics, San Diego, Calif.), which detects both GDH (TR-GDH) and toxin A (TR-Tox-A); an in-house cytotoxin assay (C-Tox); and stool cultures for C. difficile.All nonformed stool specimens from inpatients suspected of having CDAD were included in the study. Except for stool cultures, all tests were performed within 48 h of the arrival of specimens in the laboratory. Specimens were kept at 4°C if not processed immediately. A portion of stool was stored at Ϫ70°C for subsequent culture. Specimens of insufficient quantity were excluded from all tests.The results of TL-GDH and Tox-A/B were read by using a dual-wavelength spectrometer (450 and 620 nm). TR-GDH, TR-Tox-A, TL-GDH, and Tox-A/B were carried out according to the manufacturer's instruction. Details of the Triage panels and Tox-A/B and C-Tox have been described previously (16).After thawing, fecal samples were planted onto prereduced cycloserine-cefoxitin-fructose agar (cefoxitin, 8 mg/liter; cycloserine, 250 mg/liter; fructose agar; Oxoid, Ottawa, Canada) and incubated for 96 h at 35°C in an anaerobic glove box. Suspected colonies were identified based on their growth on selective media, their colonial morphology, and a positive reaction by the MicroScreen C. difficile latex slide agglutination test (Microgen Bioproducts Ltd., Surrey, United Kingdom) (2, 11).All C. difficile isolates were tested by PCR for the presence of toxin A and B genes by use of primers NK9 and NK11, derived from the repeating portion of toxin A, and NK104 and NK105, derived from the nonrepeating portion of toxin B. These primers have been described by Kato and coworkers (13). The composition of our master mix and the amplification conditions were the same as used by those investigators.For the purpose of this study, a true positive sample was defined as a sample that was positive for toxin A and/or B and from which a toxigenic C. difficile strain (positive by PCR for a toxin gene) was isolated. If a positive toxin detection test result was seen for a sample which was negative for C. difficile by culture or yielded a nontoxigenic strain, the reaction was considered to be falsely positive. Sensitivity, specificity, and positive and negative predictive values for each test were calculated on this basis.C. difficile was isolated from 93 of 49...

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supporting

confidence: 90%

“…TR-GDH, TR-Tox-A, TL-GDH, and Tox-A/B were carried out according to the manufacturer's instruction. Details of the Triage panels and Tox-A/B and C-Tox have been described previously (16).…”

mentioning

confidence: 99%

Performance of the TechLab C. DIFF CHEK-60 Enzyme Immunoassay (EIA) in Combination with the C. difficile Tox A/B II EIA Kit, the Triage C. difficile Panel Immunoassay, and a Cytotoxin Assay for Diagnosis of Clostridium difficile -Associated Diarrhea

et al. 2004

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“…Because the results for all negative specimens were not confirmed by the CTN assay, the specificity and the negative predictive value of this GDH assay cannot be stated with certainty. However, 293 of 298 of the GDH assay-negative specimens were also negative by the two other immunoassays that were studied, with the A/B EIA having been shown to be highly specific and to have a high negative predictive value in several studies (9,11,12,15,19). The seven GDH assay-negative specimens that were positive by one of the two other EIAs were found to be negative by the CTN assay.…”

mentioning

confidence: 95%

“…The latter is the most widely used test for the detection of C. difficile toxins in the United States (4). Recent studies (11,20) have shown that this assay has both excellent sensitivity (Ͼ90%) and excellent specificity (Ͼ95%) compared to the results of a tissue culture cytotoxicity neutralization (CTN) assay, an assay frequently considered the "gold standard" for the diagnosis of C. difficile disease (1,9,11,12,15,18,19,21). Other data suggest that the detection of toxigenic strains of C. difficile is actually a more sensitive approach to the diagnosis of C. difficile disease (5,6,13,14), but culture is infrequently done for diagnostic purposes in the United States because of its slow turnaround time and the need for special isolation media and technical expertise.…”

mentioning

confidence: 99%

Is a Two-Step Glutamate Dehyrogenase Antigen-Cytotoxicity Neutralization Assay Algorithm Superior to the Premier Toxin A and B Enzyme Immunoassay for Laboratory Detection of Clostridium difficile ?

Gilligan

2008

J Clin Microbiol

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A two-step algorithm for the detection of Clostridium difficile by the use of C. Diff Quik Chek (TechLab, Blacksburg, VA) and a tissue culture cytotoxicity neutralization assay was found to be more sensitive than the widely used solid-phase enzyme immunoassay (EIA), the Premier toxin A and B EIA (Meridian Bioscience, Cincinnati, OH), and a newly developed, rapid single-test EIA for C. difficile toxins A and B (Tox A/B Quik Chek; TechLab).The recent emergence of a highly virulent strain of Clostridium difficile, with its associated increased morbidity and increased mortality, has placed a renewed emphasis on the laboratory diagnosis of C. difficile disease (7, 10). For well over a decade, first the Premier toxin A enzyme immunoassay (EIA) and then the Premier toxin A and B EIA (A/B EIA; Meridian Bioscience, Inc., Cincinnati, OH) were used to detect this toxin (21). The latter is the most widely used test for the detection of C. difficile toxins in the United States (4). Recent studies (11,20) have shown that this assay has both excellent sensitivity (Ͼ90%) and excellent specificity (Ͼ95%) compared to the results of a tissue culture cytotoxicity neutralization (CTN) assay, an assay frequently considered the "gold standard" for the diagnosis of C. difficile disease (1,9,11,12,15,18,19,21). Other data suggest that the detection of toxigenic strains of C. difficile is actually a more sensitive approach to the diagnosis of C. difficile disease (5,6,13,14), but culture is infrequently done for diagnostic purposes in the United States because of its slow turnaround time and the need for special isolation media and technical expertise. In addition, the specificity of toxigenic culture for the diagnosis of C. difficile disease has also been questioned (1,14).Given the emergence of a highly virulent C. difficile strain in the United States, a recent report by Ticehurst and colleagues was a cause for concern (18). They showed that another widely used EIA, the C. difficile Tox A/B II assay (TechLab, Blacksburg, VA), which has performance characteristics similar to those of the A/B EIA (11,12,19), had a sensitivity of only 36% for C. difficile detection when the results were compared to those obtained by the use of a two-step algorithm in which specimens were first tested for the presence of the glutamate dehydrogenase (GDH) antigen by a solid-phase EIA (C. Diff Chek-60; TechLab) and the positive results were confirmed by the CTN assay. The test for the GDH antigen proved to be an excellent screening test, with a sensitivity of Ͼ95% and a negative predictive value of Ͼ99% compared with the results of the CTN assay (18). In practical terms, that meant that GDHnegative specimens could be reported to be negative without further testing. By using the CTN assay as a reference method, it was found that the assay for the GDH antigen had a positive predictive value of 53%, indicating that a confirmatory test was required to detect specimens containing C. difficile toxins.I applied the two-step algorithm approach using a modification ...

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“…The sensitivity of ELISA is 63% to 92%, and false-negative results are possible. 36 A second ELISA targets glutamate dehydrogenase (GDH), a common C difficile antigen that is secreted into the stool. 34 GDH is an enzyme present in microbes that converts glutamate to -ketoglutarate and vice versa.…”

Section: Diagnosismentioning

confidence: 99%

Clostridium difficile Infection: Clinical Challenges and Management Strategies

Walters

Zuckerbraun

2014

Critical Care Nurse

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C lostridium difficile, an anaerobic, spore-forming, toxin-forming, gram-positive bacillus, has become the leading cause of health care-associated infectious diarrhea.1,2 The spectrum of C difficile infection (CDI) ranges from uncomplicated diarrhea to systemic toxic effects marked by sepsis and death.2 Fulminant cases, requiring colectomy, have a mortality rate up to 80%. 3 CDI is increasingly common and causes marked morbidity and mortality, prolonged hospital lengths of stay, and increased health care costs. Recent excess health care costs of hospital-onset CDI are estimated to be $5042 to $7179 per case, with a national annual estimate (limited to hospital-onset CDI) of $897 million to $1.3 billion. 4 The emergence of a hypervirulent epidemic strain and community-acquired disease in populations previously at low risk for CDI most likely will place an even greater burden on patients and the health care system.

show abstract

Clinical Usefulness of Components of the Triage Immunoassay, Enzyme Immunoassay for Toxins A and B, and Cytotoxin B Tissue Culture Assay for the Diagnosis of Clostridium difficile Diarrhea

Cited by 29 publications

References 22 publications

Performance of the TechLab C. DIFF CHEK-60 Enzyme Immunoassay (EIA) in Combination with the C. difficile Tox A/B II EIA Kit, the Triage C. difficile Panel Immunoassay, and a Cytotoxin Assay for Diagnosis of Clostridium difficile -Associated Diarrhea

Performance of the TechLab C. DIFF CHEK-60 Enzyme Immunoassay (EIA) in Combination with the C. difficile Tox A/B II EIA Kit, the Triage C. difficile Panel Immunoassay, and a Cytotoxin Assay for Diagnosis of Clostridium difficile -Associated Diarrhea

Is a Two-Step Glutamate Dehyrogenase Antigen-Cytotoxicity Neutralization Assay Algorithm Superior to the Premier Toxin A and B Enzyme Immunoassay for Laboratory Detection of Clostridium difficile ?

Clostridium difficile Infection: Clinical Challenges and Management Strategies

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