Detection of Toxigenic Clostridium difficile in Stool Samples by Real-Time Polymerase Chain Reaction for the Diagnosis of C. difficile-Associated Diarrhea

Peterson, Lance R.; Manson, Rebecca U.; Paule, Suzanne M.; Hacek, Donna M.; Robicsek, Ari; Thomson, Richard B.; Kaul, Karen L.

doi:10.1086/522185

Cited by 199 publications

(176 citation statements)

References 42 publications

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“…Many studies have developed different real-time PCR (RT-PCR) methods for the detection of C. difficile toxin genes directly from stool samples, not only from humans [166,167] but also from animals [168], and for the quantitative detection of C. difficile in hospital environmental samples [169]. Various automated RT-PCR systems are commercially available, intended as diagnostic tools for CDI.…”

Section: Laboratory Diagnosis Of CDImentioning

confidence: 99%

Clostridium difficile infection: Early history, diagnosis and molecular strain typing methods

Rodríguez

Broeck

Taminiau

et al. 2016

Microbial Pathogenesis

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a b s t r a c tRecognised as the leading cause of nosocomial antibiotic-associated diarrhoea, the incidence of Clostridium difficile infection (CDI) remains high despite efforts to improve prevention and reduce the spread of the bacterium in healthcare settings. In the last decade, many studies have focused on the epidemiology and rapid diagnosis of CDI. In addition, different typing methods have been developed for epidemiological studies. This review explores the history of C. difficile and the current scope of the infection. The variety of available laboratory tests for CDI diagnosis and strain typing methods are also examined.

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Section: Laboratory Diagnosis Of CDImentioning

confidence: 99%

Clostridium difficile infection: Early history, diagnosis and molecular strain typing methods

Rodríguez

Broeck

Taminiau

et al. 2016

Microbial Pathogenesis

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“…The increase in the number of cases cannot be explained solely by the difference in performance characteristics of the 2 tests: the sensitivity (68%-90% for EIA vs 88%-100% for PCR) and specificity (95.3%-99% for EIA vs 92.6%-98.4% for PCR) of the 2 tests are comparable. [12][13][14]26,27 The sudden increase in the incidence of CDI following the implementation of PCR could be partly explained by the fact that PCR detects toxin genes but cannot determine whether the organism is actively producing toxin. For that reason, some studies concluded that PCR is unreliable in differentiating CDI cases from asymptomatic carriers of a potentially toxigenic organism.…”

Section: Discussionmentioning

confidence: 99%

“…10,11 Several studies showed that PCR had an equivalent sensitivity and specificity (up to 100%) compared with toxigenic cultures. [12][13][14] However, PCR may not be useful when trying to differentiate carrier status from true CDI. 15 Moreover, given that clinical manifestations of CDI are milder in children compared with the adult population, 16 and given the current absence of testing strategies that accurately and optimally diagnose CDI, 17 we aimed to determine the proportion of pediatric patients diagnosed as having CDI by PCR who would also be diagnosed by EIA, and to compare the clinical characteristics of PCR + /enzyme-linked immunosorbent assay (ELISA) + vs PCR + /ELISA − patients.…”

mentioning

confidence: 99%

Clostridium difficile Infections in Children: Impact of the Diagnostic Method on Infection Rates

Alghounaim

Longtin

Gonzales

et al. 2016

Infect. Control Hosp. Epidemiol.

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BACKGROUNDPolymerase chain reaction (PCR) assays based on the detection of the toxin B gene are replacing enzyme-linked immunosorbent assay (ELISA)–based toxin production detection or cell cytotoxicity assay in most laboratories.OBJECTIVETo determine the proportion of pediatric patients diagnosed withClostridium difficile infection by PCR who would have also been diagnosed by ELISA and to compare the clinical characteristics of PCR+/ELISA+ vs PCR+/ELISA− patients.METHODSUsing the microbiology laboratory information system, stool samples positive for C. difficile by PCR between October 2010 and July 2014 were identified. Using frozen stool specimens, an ELISA for toxin A and B was performed. A retrospective medical chart review was conducted to obtain demographic and clinical data. Duplicate samples were excluded.RESULTSA total of 136 PCR-positive samples underwent ELISA testing: 54 (40%) were positive for toxin A or B. The mean (SD) age of the entire cohort was 8.5 (6.2) years. There was no difference in age, gender, clinical manifestation, previous medical problems, and management between patients positive or negative by ELISA. However, patients positive by ELISA were more likely to have had a recent exposure to antibiotics (67.9% vs 50%; crude odds ratio, 2.1 [95% CI, 1.03–4.28]).CONCLUSIONIn our pediatric population, 60% of patients with C. difficile diagnosed by PCR had no toxin detectable by ELISA. ELISA-negative patients were less likely to have received an antibiotic recently compared with ELISA-positive patients. These results highlight the need to standardize laboratory criteria for the diagnosis of C. difficile infections in children.Infect Control Hosp Epidemiol 2016;37:1087–1093

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“…Foremost among the findings was the absence of any documentation of diarrhea in more than one-third of patients whose records were accessible. In their prospective study, Peterson and colleagues 15 found a similarly high proportion (39%) of potential CDI episodes that did not meet criteria for diarrhea (ie, $3 loose stools in 1 day). The primary limitation of the current approach was its retrospective nature, inasmuch as the absence of documented diarrhea cannot be interpreted definitively as a documented absence; yet the lack of mention altogether in such a large proportion of cases is noteworthy given the fact that diarrhea is the most common underlying feature of CDI.…”

Section: Commentmentioning

confidence: 92%

“…14, 15 The latter approach is hampered by successive decreases in PPV with each sequential test, as illustrated by Peterson and Robicsek. 16 Improvement of immunoassay formats that detect C difficile-specific GDH marked a potential diagnostic advancement with minimal increases in assay cost compared to that of toxin EIAs.…”

Section: Commentmentioning

confidence: 99%

Comparison of Analytical and Clinical Performance of Three Methods for Detection of Clostridium difficile

LaSala

Svensson²,

Mohammad³

et al. 2012

Archives of Pathology &Amp; Laboratory Medicine

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difficile infection has undergone considerable and rapid evolution during the last decade. The ideal detection method(s), which should exhibit high analytical and clinical sensitivity and specificity, remains undefined.Objective.-We sought to evaluate the analytical and clinical performance characteristics of three methods for the laboratory detection of C difficile.Design.-This study used 114 consecutive stool samples to compare three methods of C difficile detection: an enzyme immunoassay (EIA) for toxins A/B, a lateral flow membrane immunoassay for glutamate dehydrogenase (GDH), and a qualitative real-time polymerase chain reaction (PCR) assay. Medical records of all patients having $1 positive test result were reviewed to estimate the clinical likelihood of C difficile infection.Results.-Based upon laboratory result consensus values, analytical sensitivity was significantly higher for GDH (94%) and PCR (94%) assays than for toxin EIA (25%). Analytical specificity was significantly higher for PCR (100%) and EIA (100%) than for GDH assay (93%). In contrast, assay performance based upon clinical probability of C difficile infection suggested lower discriminatory power (ie, clinical specificity) of the more analytically sensitive methods.Conclusions.-Higher rates of C difficile detection will be realized upon implementation of GDH assay and/or real-time PCR-based testing algorithms than by testing with EIA alone. Further study is required to elucidate potential downstream costs for higher detection rates.

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Detection of Toxigenic Clostridium difficile in Stool Samples by Real-Time Polymerase Chain Reaction for the Diagnosis of C. difficile-Associated Diarrhea

Cited by 199 publications

References 42 publications

Clostridium difficile infection: Early history, diagnosis and molecular strain typing methods

Clostridium difficile infection: Early history, diagnosis and molecular strain typing methods

Clostridium difficile Infections in Children: Impact of the Diagnostic Method on Infection Rates

Comparison of Analytical and Clinical Performance of Three Methods for Detection of Clostridium difficile

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