Validation of Cepheid Xpert® C. difficile assay using 400µL liquid transport medium from Copan Fecal Swab® for the detection of Clostridium difficile DNA in stool samples positive for glutamate dehydrogenase antigen

2016 ◽  
Author(s):  
Yvan Caspar
Keyword(s):  
Clostridium Difficile ◽  
Glutamate Dehydrogenase ◽  
Liquid Transport ◽  
Transport Medium ◽  
Stool Samples

scholarly journals Clostridium difficile glutamate dehydrogenase is a secreted enzyme that confers resistance to H2O2

Microbiology ◽  
2014 ◽  
Vol 160 (1) ◽  
pp. 47-55 ◽  
Author(s):  
Brintha Prasummanna Girinathan ◽  
Sterling E. Braun ◽  
Revathi Govind
Keyword(s):  
Clostridium Difficile ◽  
Glutamate Dehydrogenase ◽  
Parent Strain ◽  
Diagnostic Tools ◽  
Growth Defect ◽  
Irreversible Reaction ◽  
Yeast Extract Medium ◽  
Stool Samples ◽  
Higher Sensitivity

Clostridium difficile produces an NAD-specific glutamate dehydrogenase (GDH), which converts l-glutamate into α-ketoglutarate through an irreversible reaction. The enzyme GDH is detected in the stool samples of patients with C. difficile‐associated disease and serves as one of the diagnostic tools to detect C. difficile infection (CDI). We demonstrate here that supernatant fluids of C. difficile cultures contain GDH. To understand the role of GDH in the physiology of C. difficile, an isogenic insertional mutant of gluD was created in strain JIR8094. The mutant failed to produce and secrete GDH as shown by Western blot analysis. Various phenotypic assays were performed to understand the importance of GDH in C. difficile physiology. In TY (tryptose yeast extract) medium, the gluD mutant grew slower than the parent strain. Complementation of the gluD mutant with the functional gluD gene reversed the growth defect in TY medium. The presence of extracellular GDH may have a functional role in the pathogenesis of CDI. In support of this assumption we found higher sensitivity to H2O2 in the gluD mutant as compared to the parent strain. Complementation of the gluD mutant with the functional gluD gene reversed the H2O2 sensitivity.

Evaluation of the new CE-IVD marked BD MAX Cdiff Assay for the detection of toxigenic Clostridium difficile harboring the tcdB gene from clinical stool samples

2013 ◽  
Vol 94 (1) ◽  
pp. 58-60 ◽  
Author(s):  
Paul O. Verhoeven ◽  
Anne Carricajo ◽  
Sylvie Pillet ◽  
Alain Ros ◽  
Nathalie Fonsale ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Stool Samples

scholarly journals Complete Microbiota Engraftment Is Not Essential for Recovery from Recurrent Clostridium difficile Infection following Fecal Microbiota Transplantation

mBio ◽  
2016 ◽  
Vol 7 (6) ◽  
Author(s):  
Christopher Staley ◽  
Colleen R. Kelly ◽  
Lawrence J. Brandt ◽  
Alexander Khoruts ◽  
Michael J. Sadowsky
Keyword(s):  
Clostridium Difficile ◽  
Bacterial Communities ◽  
Healthy Donor ◽  
Fecal Microbiota Transplantation ◽  
Fecal Microbiota ◽  
Mean Value ◽  
Bile Acid Metabolism ◽  
Secondary Bile Acid ◽  
Stool Samples ◽  
Relative Abundances

ABSTRACT Bacterial communities from subjects treated for recurrent Clostridium difficile infection (rCDI) by fecal microbiota transplantation (FMT), using either heterologous donor stool samples or autologous stool samples, were characterized by Illumina next-generation sequencing. As previously reported, the success of heterologous FMT (90%) was superior to that of autologous FMT (43%) ( P = 0.019), and post-FMT intestinal bacterial communities differed significantly between treatment arms ( P < 0.001). Subjects cured by autologous FMT typically had greater abundances of the Clostridium XIVa clade and Holdemania bacteria prior to treatment, and the relative abundances of these groups increased significantly after FMT compared to heterologous FMT and pre-FMT samples. The typical shift to post-FMT, donor-like assemblages, featuring high relative abundances of genera within the Bacteroidetes and Firmicutes phyla, was not observed in the autologous FMT subjects. Autologous FMT patient bacterial communities were significantly different in composition than those for heterologous FMT patients and donors ( P < 0.001). The SourceTracker program, which employs a Bayesian algorithm to determine source contributions to sink communities, showed that patients initially treated by heterologous FMT had significantly higher percentages of engraftment (i.e., similarity to donor communities, mean value of 74%) compared to those who suffered recurrence following autologous FMT (1%) ( P ≤ 0.013). The findings of this study suggest that complete donor engraftment may be not necessary if functionally critical taxa are present in subjects following antibiotic therapy. IMPORTANCE This study provides a detailed characterization of fecal bacterial communities in subjects who participated in a previously published randomized clinical trial to treat recurrent C. difficile infection (rCDI). Bacterial communities were characterized to determine differences between subjects who received fecal bacteria either from healthy donor stool samples or their own stool samples as “placebo” in order to determine which groups of bacteria were most important in achieving a cure. The results of this study suggested that bacteria associated with secondary bile acid metabolism could potentially provide resistance to infection and that complete transfer of healthy donor microorganisms was not necessary to resolve CDI following unsuccessful antibiotic treatment.

scholarly journals Multiplex Real-Time PCR Method for Simultaneous Identification and Toxigenic Type Characterization of Clostridium difficile From Stool Samples

2015 ◽  
Vol 35 (3) ◽  
pp. 306-313 ◽  
Author(s):  
Abdullah Kilic ◽  
Mohammad J. Alam ◽  
Naradah L. Tisdel ◽  
Dhara N. Shah ◽  
Mehmet Yapar ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Real Time ◽  
Real Time Pcr ◽  
Stool Samples ◽  
Pcr Method ◽  
Simultaneous Identification

scholarly journals Detection of Clostridium difficile Infection in Al-Quwayiyah General Hospital, Riyadh, Kingdom of Saudi Arabia

2020 ◽  
pp. 94-102
Author(s):  
Enas Sh. Khater ◽  
Abd Alazim A. Al- Faki
Keyword(s):  
Saudi Arabia ◽  
Abdominal Pain ◽  
Clostridium Difficile ◽  
Proton Pump ◽  
Peptic Ulcers ◽  
Pcr Assay ◽  
Kingdom Of Saudi Arabia ◽  
Statistical Significant Difference ◽  
Significant Difference ◽  
Stool Samples

Clostridium difficile infections (CDIs) is considered healthcare-associated infections which cause watery diarrhea to long stayed hospitalized patients and cause increased mortality rate. Aim: Detection of the prevalence and risk factors of C. difficile in Al Quwayiyah General hospital, Riyadh, Kingdom of Saudi Arabia and compairing between GeneXpert® PCR assay and Quikchek complete-enzyme imunoassay QCC, (QCC-EIA) in detection of C. difficile infection and toxicity Materials and Methods: A cross sectional and prospective study was performed for one year started from June 2019 to June 2020. The data collected include demographic, laboratory and clinical data. A total of 104 stool samples were collected from patients presented with diarrhea. GeneXpert® PCR assay and Quikchek complete-enzyme imunoassay QCC (QCC-EIA) were conducted to each stool sample. Results: Only 15(14.4%) of the 104 studied patients had CDI while 89 (85.6%) were non CDI patients, 13 (86.7%) of the CDI patients were males and 2 (13.3%) were females with mean age for CDI cases 61 (±19.9), while non CDI cases involved 55(61.8%) were males and 34 (38.2%) were females with mean age for cases of non CDI, 60 (±18.7) years. Of the CDI and non CDI cases respectively 12 (80%) and 14(15.7%) had fever, 5 (27%) and 6 (6.7%) had vomitting and 7 (46.7%) and 12 (13.5%) of cases had abdominal pain. There was statistical significant difference between patients with fever while no statistical significant difference regarding vomitting and abdominal pain. There was statistical significant difference between patients with peptic ulcers, patients received proton pump inhibitors and patients received broad-spectrum antibiotics, while There was no statistical significant difference between cardiac disease, cerebrovascular disease, diabetes, pulmonary disease, hepatic disease and Renal disease. Gene expert PCR detected 15/104(14.4%) as positive CDI while QCC-EIA detected 21/104 (20.5%) as positive CDI. On comparison between gene expert PCR technique and QCC-EIA the sensitivity of QCC-EIA was 100%, while the specificity was 91%. The Positive Predictive Value was 74%, while the Negative Predictive Value was 100%. Conclusion: The C. difficile infection prevalence rate in the hospital was 14.4%. There was statistical significant difference between patients with peptic ulcers, patients received proton pump inhibitors and patients received broad-spectrum antibiotics. The QCC-EIA can be used as a screening test for the detection of C. difficile toxin in stool samples but should be confirmed with a PCR assay or another confirmatory test Due to its decreased specificity.

scholarly journals Clostridium difficile colonization among patients with clinically significant diarrhea and no identifiable cause of diarrhea

2018 ◽  
Vol 39 (11) ◽  
pp. 1330-1333 ◽  
Author(s):  
Erik R. Dubberke ◽  
Kimberly A. Reske ◽  
Tiffany Hink ◽  
Jennie H. Kwon ◽  
Candice Cass ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Patient Population ◽  
Tertiary Care ◽  
Clinical Microbiology Laboratory ◽  
Care Hospital ◽  
Toxigenic Culture ◽  
Stool Samples ◽  
Stool Specimens ◽  
Clinically Significant ◽  
Culture Negative

AbstractObjectiveTo determine the prevalence of Clostridium difficile colonization among patients who meet the 2017 IDSA/SHEA C. difficile infection (CDI) Clinical Guideline Update criteria for the preferred patient population for C. difficile testing.DesignRetrospective cohort.SettingTertiary-care hospital in St. Louis, Missouri.PatientsPatients whose diarrheal stool samples were submitted to the hospital’s clinical microbiology laboratory for C. difficile testing (toxin EIA) from August 2014 to September 2016.InterventionsElectronic and manual chart review were used to determine whether patients tested for C. difficile toxin had clinically significant diarrhea and/or any alternate cause for diarrhea. Toxigenic C. difficile culture was performed on all stool specimens from patients with clinically significant diarrhea and no known alternate cause for their diarrhea.ResultsA total of 8,931 patients with stool specimens submitted were evaluated: 570 stool specimens were EIA positive (+) and 8,361 stool specimens were EIA negative (−). Among the EIA+stool specimens, 107 (19% of total) were deemed eligible for culture. Among the EIA− stool specimens, 515 (6%) were eligible for culture. One EIA+stool specimen (1%) was toxigenic culture negative. Among the EIA− stool specimens that underwent culture, toxigenic C. difficile was isolated from 63 (12%).ConclusionsMost patients tested for C. difficile do not have clinically significant diarrhea and/or potential alternate causes for diarrhea. The prevalence of toxigenic C. difficile colonization among EIA− patients who met the IDSA/SHEA CDI guideline criteria for preferred patient population for C. difficile testing was 12%.

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