Development of a high-sensitivity nested PCR assay for the detection of Clostridium piliforme in clinical samples

Tyzzer’s disease has been reported in numerous animal species, including rodents and lagomorphs such as the rabbit. Because the organism cannot be cultivated on artificial media the diagnosis of the aetiological agent Clostridium piliforme is rather difficult and is based on the identification of typical gross lesions and histological demonstration of the characteristic intracellular bacteria at the periphery of the necrotic foci. To address the limitations of C. piliforme detection a nested polymerase chain reaction (nPCR) assay has been developed with increased specificity with regard to the phylogenetically close bacterial strains (based on 16S RNA sequence analogies). The sensitivity of this nPCR was also demonstrated using clinical samples.