Evaluation of the usefulness of counterimmunoelectrophoresis for diagnosis of Clostridium difficile-associated colitis in clinical specimens

Wu, Tina; Fung, J C

doi:10.1128/jcm.17.4.610-613.1983

Cited by 25 publications

(11 citation statements)

References 15 publications

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“…42 The increased abundance of members of the Clostridium genus observed here was not unexpected and might reflect their ability to form spores that are highly resilient to antibiotics. Indeed, reports of increasing C. difficile carriage rates [43][44][45][46][47] possibly reflect increases in cumulative antibiotic exposure that accompany improved CF patient longevity. 48 In contrast, commensal members of the Clostridiales order that are not capable of sporulation, such as the Faecalibacterium and Ruminococcus genera, showed reduced relative abundance compared to healthy controls.…”

Section: Discussionmentioning

confidence: 99%

Opportunistic bacteria confer the ability to ferment prebiotic starch in the adult cystic fibrosis gut

et al. 2018

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Chronic disruption of the intestinal microbiota in adult cystic fibrosis (CF) patients is associated with local and systemic inflammation, and has been linked to the risk of serious comorbidities. Supplementation with high amylose maize starch (HAMS) might provide clinical benefit by promoting commensal bacteria and the biosynthesis of immunomodulatory metabolites. However, whether the disrupted CF gut microbiota has the capacity to utilise these substrates is not known. We combined metagenomic sequencing, in vitro fermentation, amplicon sequencing, and metabolomics to define the characteristics of the faecal microbiota in adult CF patients and assess HAMS fermentation capacity. Compared to healthy controls, the faecal metagenome of adult CF patients had reduced bacterial diversity and prevalence of commensal fermentative clades. In vitro fermentation models seeded with CF faecal slurries exhibited reduced acetate levels compared to healthy control reactions, but comparable levels of butyrate and propionate. While the commensal genus Faecalibacterium was strongly associated with short chain fatty acid (SCFA) production by healthy microbiota, it was displaced in this role by Clostridium sensu stricto 1 in the microbiota of CF patients. A subset of CF reactions exhibited enterococcal overgrowth, resulting in lactate accumulation and reduced SCFA biosynthesis. The addition of healthy microbiota to CF faecal slurries failed to displace predominant CF taxa, or substantially influence metabolite biosynthesis. Despite significant microbiota disruption, the adult CF gut microbiota retains the capacity to exploit HAMS. Our findings highlight the potential for taxa associated with the altered CF gut microbiotato mediate prebiotic effects in microbial systems subject to ongoing perturbation, irrespective of the depletion of common commensal clades. ARTICLE HISTORY

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Section: Discussionmentioning

confidence: 99%

Opportunistic bacteria confer the ability to ferment prebiotic starch in the adult cystic fibrosis gut

et al. 2018

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“…The growing concern among physicians regarding enteric diseases associated with C. difficile has placed a heavy demand on clinical microbiology laboratories to offer a rapid and reliable diagnostic test. Methods proposed so far include counterimmunoelectrophoresis (26,35), latex agglutination (15,16,20), dot immunobinding (34), and EIAs using polyclonal antiserum (1,18,19,31), but all lack either sensitivity or specificity. The stool cytotoxicity assay is still considered the gold standard for C. difficile toxin B in stool specimens because of its high sensitivity (<1 pg of toxin B) and specificity (neutralization of the cytopathic effect by a specific antiserum) (7).…”

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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“…The search for rapid techniques has evolved from counterimmunoelectrophoresis assays developed in the early 1980's, which suffered from lack of both specificity and sensitivity (18,20,22,24), to LA and more recently to EIA methods, which have become commercially available and are gaining popularity because of their rapidity and ease of performance.…”

Section: Discussionmentioning

confidence: 99%

Multicenter evaluation of a new enzyme immunoassay for detection of Clostridium difficile enterotoxin A

et al. 1992

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The Premier Clostridium diffwile toxin A enzyme immunoassay (PTA ETA) (Meridian Diagnostics, Inc., Cincinnati, Ohio) for rapid diagnosis of antibiotic-associated colitis (AAC) was evaluated in a multicenter study. Stool samples from 421 patients suspected of having AAC were tested for toxin A by the PTA EIA and for toxin B by three tissue culture assays (TCA) employing WI-38 cells (New England Deaconess Hospital) in conventional tubes or foreskin fibroblasts (Children's Hospital) or Vero cells (Beth Israel Hospital) in microwells. The tubes and plates were examined at 24 and 48 h for cytotoxicity. Clinical criteria, repeat testing at another site, and culture of frozen stool samples for C. diJficik were used to evaluate discrepant results. Of 504 samples, 66 were positive and 409 were negative by both tests. Eight samples had indeterminate PTA EIA results and were excluded from this analysis. Of 21 discrepancies, 9 were PTA EIA positive and TCA negative and 12 were PTA EIA negative TCA positive. Following resolution of the discrepancies, 11 of 12 PTA EIA-negative-TCA-positive and 5 of 9 PTA EIA-positive-TCA-negative samples were considered true positive for AAC. The sensitivity and specificity were, respectively, 86.6 and 99.0% for the PTA EIA and 93.9 and 99.8% for TCA. The predictive values of positive and negative tests were, respectively, 94.7 and 97.4% for the PTA EIA and 98.7 and 98.8% for TCA. We conclude that the PTA EIA is a rapid, simple EIA technique whose accuracy in detecting enterotoxin A approaches that of reference TCA methods for detection of cytotoxin B.

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Evaluation of the usefulness of counterimmunoelectrophoresis for diagnosis of Clostridium difficile-associated colitis in clinical specimens

Cited by 25 publications

References 15 publications

Opportunistic bacteria confer the ability to ferment prebiotic starch in the adult cystic fibrosis gut

Opportunistic bacteria confer the ability to ferment prebiotic starch in the adult cystic fibrosis gut

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

Multicenter evaluation of a new enzyme immunoassay for detection of Clostridium difficile enterotoxin A

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