Quantification of Clostridium difficile in Antibiotic-Associated-Diarrhea Patients

Naaber, Paul; Štšepetova, Jelena; Smidt, Imbi; Rätsep, Merle; Kõljalg, Siiri; Lõivukene, Krista; Jaanimäe, Liis; Löhr, Iren Høyland; Natås, Olav B.; Truusalu, Kai; Sepp, Epp

doi:10.1128/jcm.05115-11

Cited by 31 publications

(32 citation statements)

References 16 publications

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“…Our study provides evidence of an association between C. difficile fecal load and the results of different diagnostic tests used routinely by clinical laboratories. Our results are in line with another recently published study showing that ToxAB-positive samples have higher bacterial loads than ToxABnegative samples (7.0 versus Ͻ2 log 10 CFU/g) (18).…”

Section: Discussionsupporting

confidence: 83%

“…Other studies support the notion that there is a significant correlation between results obtained by real-time PCR and quantitative culture (18,30). This suggests that a readily available qualitative real-time PCR that does not require calibration can provide some approximate indication of the C. difficile fecal load.…”

Section: Discussionsupporting

confidence: 57%

“…We developed a quantitative culture protocol adapted from previous studies (18)(19)(20)(21). Briefly, a 0.15-g aliquot of the sample was transferred to a conical tube and treated by ethanol shock for 30 min in an equal volume of 90% ethanol.…”

Section: Methodsmentioning

confidence: 99%

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Correlation between Clostridium difficile Bacterial Load, Commercial Real-Time PCR Cycle Thresholds, and Results of Diagnostic Tests Based on Enzyme Immunoassay and Cell Culture Cytotoxicity Assay

et al. 2013

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The impact of Clostridium difficile fecal loads on diagnostic test results is poorly understood, but it may have clinical importance. In this study, we investigated the relationship between C. difficile fecal load and the results of four assays: a glutamate dehydrogenase (GDH) enzyme immunoassay (EIA), a toxin A/B antigen EIA (ToxAB), a cell culture cytotoxicity assay (CCA), and PCR targeting the tcdB gene. We also compared the PCR cycle threshold (C T ) with the results of quantitative culture using Spearman's rank correlation coefficient. .35 to 7.83), respectively. Group 1 samples had lower fecal loads than those from each of the other groups (P < 0.001). Group 2 samples had lower fecal loads than those from groups 3 and 4 (P < 0.001). There was a significant correlation between PCR C T and fecal loads ( ‫؍‬ ؊0.697; P < 0.001). NAP1 strains were associated with the detection of toxins by EIA or CCA (P ‫؍‬ 0.041). This study demonstrates an association between C. difficile fecal load and the results of routinely used diagnostic tests.

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Section: Discussionsupporting

confidence: 83%

Section: Discussionsupporting

confidence: 57%

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Correlation between Clostridium difficile Bacterial Load, Commercial Real-Time PCR Cycle Thresholds, and Results of Diagnostic Tests Based on Enzyme Immunoassay and Cell Culture Cytotoxicity Assay

et al. 2013

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“…It is notable that the mean stool density of C. difficile among our healthy participants was 3.0 ϫ 10 4 CFU/g, whereas the reported value for C. difficile stool density in symptomatic CDI patients was reported to be 4.0 ϫ 10 6 CFU/g (14). C. difficile-colonized individuals with a low colonic density of organisms may not participate in the CDI transmission cycle; this is unlikely, however, because the infectious doses for CDIs in animal models are low (15) and C. difficile-colonized patients within hospitals are well documented to be responsible for contamination of hospital rooms and transmission to new patients (16)(17)(18).…”

Section: Discussionmentioning

confidence: 49%

Prevalence and Duration of Asymptomatic Clostridium difficile Carriage among Healthy Subjects in Pittsburgh, Pennsylvania

et al. 2014

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bPrevious studies suggested that 7 to 15% of healthy adults are colonized with toxigenic Clostridium difficile. To investigate the epidemiology, genetic diversity, and duration of C. difficile colonization in asymptomatic persons, we recruited healthy adults from the general population in Allegheny County, Pennsylvania. Participants provided epidemiological and dietary intake data and submitted stool specimens. The presence of C. difficile in stool specimens was determined by anaerobic culture. Stool specimens yielding C. difficile underwent nucleic acid testing of the tcdA gene segment with a commercial assay; tcdC genotyping was performed on C. difficile isolates. Subjects positive for C. difficile by toxigenic anaerobic culture were asked to submit additional specimens. One hundred six (81%) of 130 subjects submitted specimens, and 7 (6.6%) of those subjects were colonized with C. difficile. Seven distinct tcdC genotypes were observed among the 7 C. difficile-colonized individuals, including tcdC genotype 20, which has been found in uncooked ground pork in this region. Two (33%) out of 6 C. difficile-colonized subjects who submitted additional specimens tested positive for identical C. difficile strains on successive occasions, 1 month apart. The prevalence of C. difficile carriage in this healthy cohort is concordant with prior estimates. C. difficile-colonized individuals may be important reservoirs for C. difficile and may falsely test positive for infections due to C. difficile when evaluated for community-acquired diarrhea caused by other enteric pathogens.C lostridium difficile infections (CDIs) have shown dramatic increases in incidence and morbidity during the past decade (1). While C. difficile is well established as a health careassociated pathogen, incidence estimates for community-acquired CDIs vary between 6 and 30% (2-4). The source of C. difficile in community-acquired cases is unclear. Prevalence estimates for asymptomatic C. difficile colonization range from 7 to 15% among healthy, non-health care workers outside the United States to 1 to 4% among health care workers in the United States and abroad (5-9). Individuals colonized with C. difficile could represent a potential reservoir of strains imported into hospitals, as well as a source of false-positive clinical test results for CDIs among patients with community-acquired diarrheal illness, especially norovirus and Clostridium perfringens. We studied the prevalence, genotypic distribution, and duration of C. difficile carriage among healthy adults in Allegheny County, Pennsylvania, and evaluated potential risk factors for asymptomatic colonization. We also examined the relative abundance of C. difficile and the sensitivity of a commercial nucleic acid test for detection of C. difficile in the stool of healthy colonized individuals. MATERIALS AND METHODSHealthy adult residents (Ն18 years of age) in Allegheny County, Pennsylvania, were recruited for participation via print and electronic advertisements in September 2012 through April 2013. Ind...

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“…Spores are uncultivable through normal plating and culture methods; hence, copy number yield was higher than CFU input for the more efficient lysis methods. 21 The poor efficiency for Salmonella spp. suggests that their nucleic acids may be subject to effects that have failed to be identified through these experiments.…”

Section: Extraction Optimisationmentioning

confidence: 99%

Can rapid integrated polymerase chain reaction-based diagnostics for gastrointestinal pathogens improve routine hospital infection control practice? A diagnostic study

Pankhurst

Macfarlane-Smith

Buchanan

et al. 2014

Health Technology Assessment

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This journal is a member of and subscribes to the principles of the Committee on Publication Ethics (COPE) (www.publicationethics.org/).Editorial contact: nihredit@southampton.ac.ukThe full HTA archive is freely available to view online at www.journalslibrary.nihr.ac.uk/hta. Print-on-demand copies can be purchased from the report pages of the NIHR Journals Library website: www.journalslibrary.nihr.ac.uk Criteria for inclusion in the Health Technology Assessment journalReports are published in Health Technology Assessment (HTA) if (1) they have resulted from work for the HTA programme, and (2) they are of a sufficiently high scientific quality as assessed by the reviewers and editors.Reviews in Health Technology Assessment are termed 'systematic' when the account of the search appraisal and synthesis methods (to minimise biases and random errors) would, in theory, permit the replication of the review by others. HTA programmeThe HTA programme, part of the National Institute for Health Research (NIHR), was set up in 1993. It produces high-quality research information on the effectiveness, costs and broader impact of health technologies for those who use, manage and provide care in the NHS. 'Health technologies' are broadly defined as all interventions used to promote health, prevent and treat disease, and improve rehabilitation and long-term care.The journal is indexed in NHS Evidence via its abstracts included in MEDLINE and its Technology Assessment Reports inform National Institute for Health and Care Excellence (NICE) guidance. HTA research is also an important source of evidence for National Screening Committee (NSC) policy decisions.For more information about the HTA programme please visit the website: http://www.nets.nihr.ac.uk/programmes/hta This reportThe research reported in this issue of the journal was funded by the HTA programme as project number 08/13/35. The contractual start date was in April 2010. The draft report began editorial review in September 2013 and was accepted for publication in January 2014. The authors have been wholly responsible for all data collection, analysis and interpretation, and for writing up their work. The HTA editors and publisher have tried to ensure the accuracy of the authors' report and would like to thank the reviewers for their constructive comments on the draft document. However, they do not accept liability for damages or losses arising from material published in this report.This report presents independent research funded by the National Institute for Health Research (NIHR). The views and opinions expressed by authors in this publication are those of the authors and do not necessarily reflect those of the NHS, the NIHR, NETSCC, the HTA programme or the Department of Health. If there are verbatim quotations included in this publication the views and opinions expressed by the interviewees are those of the interviewees and do not necessarily reflect those of the authors, those of the NHS, the NIHR, NETSCC, the HTA programme or the Department of Health. Published by t...

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Quantification of Clostridium difficile in Antibiotic-Associated-Diarrhea Patients

Cited by 31 publications

References 16 publications

Correlation between Clostridium difficile Bacterial Load, Commercial Real-Time PCR Cycle Thresholds, and Results of Diagnostic Tests Based on Enzyme Immunoassay and Cell Culture Cytotoxicity Assay

Correlation between Clostridium difficile Bacterial Load, Commercial Real-Time PCR Cycle Thresholds, and Results of Diagnostic Tests Based on Enzyme Immunoassay and Cell Culture Cytotoxicity Assay

Prevalence and Duration of Asymptomatic Clostridium difficile Carriage among Healthy Subjects in Pittsburgh, Pennsylvania

Can rapid integrated polymerase chain reaction-based diagnostics for gastrointestinal pathogens improve routine hospital infection control practice? A diagnostic study

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