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Detection of toxigenic Clostridium difficile in fecal samples by colony blot hybridization
M J Wolfhagen1, A C Fluit, M Jansze
1Eijkman-Winkler Laboratory for Medical Microbiology, University Hospital Utrecht, The Netherlands.
Abstract:
A hybridization assay for detection of toxigenic Clostridium difficile in fecal samples was developed and compared with the classical tissue culture cytotoxicity assay. A DNA fragment probe specific for the toxin B gene of Clostridium difficile was synthesized by the polymerase chain reaction and labelled with digoxigenin. Fecal samples were cultured for 24 hours, replica-plated and hybridized with the probe. The hybridization assay had a sensitivity of 100%, specificity of 96.7%, positive predictive value of 86.7% and negative predictive value of 100% compared with the cytotoxicity assay.