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Title: Rapid detection of Clostridium difficile toxins from stool samples using real-time multiplex PCR.
Authors: Pallis, Ann;Jazayeri, Jalal;Ward, Peter;Dimovski, Karolina;Svobodova, Suzanne
Molecular Diagnostic and Microbiology Laboratory, Austin Pathology, Melbourne, VIC 3084, Australia
Issue Date: 20-Jun-2013
Citation: Journal of Medical Microbiology 2013; 62(Pt 9): 1350-6
Abstract: In this study, a total of 650 stool samples were tested to show that our method is capable of detecting four Clostridium difficile genes; tcdA, tcdB, encoding toxin A (TcdA) and toxin B (TcdB), and the binary toxin C. difficile transferase genes (cdtA and/or cdtB) encoding CDT toxin. Besides detecting the targeted C. difficile genes, our method can be used to detect the presence of any inhibitory components in the PCR. This assay, combined with a selective culture medium, such as the chromID™ C. difficile, can be applied directly for screening C. difficile-associated disease. The PCR-based assay developed here is rapid (4 h per 21 stool samples) and accurate in diagnosing C. difficile infection, 100 % assay sensitivity and negative predictive value (NPV) were obtained. However, the assay specificity of 99.1 % and positive predictive value (PPV) of 94.9 % were slightly lower than the optimal value of 100 %. The assay protocol outlined here can be used as a rapid screening tool to assist infection control units and in managing infected patients by reducing the number of patients requiring isolation and extended hospitalization. Rapid detection can prevent unnecessary antibiotic therapy and potentially reduce the spread of infection by emerging hypervirulent C. difficile strains.
Internal ID Number: 23788597
DOI: 10.1099/jmm.0.058339-0
Type: Journal Article
Subjects: Bacterial Proteins.genetics
Bacterial Toxins.genetics
Clostridium Infections.diagnosis
Clostridium difficile.genetics.isolation & purification.metabolism
Culture Media.metabolism
Genes, Bacterial
Multiplex Polymerase Chain Reaction.methods
Predictive Value of Tests
Reproducibility of Results
Sensitivity and Specificity
Appears in Collections:Journal articles

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