1 National Food Institute, Technical University of Denmark2 Division of Food Microbiology, National Food Institute, Technical University of Denmark3 French Agency for Food, Environmental and Occupational Health & Safety4 National Veterinary Institute5 National Reference Centre for Botulism6 Instituto Zooprofilattico Sperimentale delle Venezie7 Research Group for Diagnostic Engineering, National Food Institute, Technical University of Denmark8 National Institute of Public Health and the Environment9 Wageningen IMARES10 Analysis and Development Laboratory 2211 National Institute of Public Health and the Environment
Two real-time PCR arrays based on the GeneDisc® cycler platform (Pall-GeneDisc Technologies) were evaluated in a multicenter collaborative trial for their capacity to specifically detect and discriminate Clostridium botulinum types C, D and their mosaic variants C-D and D-C that are associated with avian and mammalian botulism. The GeneDisc® arrays developed as part of the DG Home funded European project 'AnibioThreat' were highly sensitive and specific when tested on pure isolates and naturally contaminated samples (mostly clinical specimen from avian origin). Results of the multicenter collaborative trial involving eight laboratories in five European Countries (two laboratories in France, Italy and The Netherlands, one laboratory in Denmark and Sweden), using DNA extracts issued from 33 pure isolates and 48 naturally contaminated samples associated with animal botulism cases, demonstrated the robustness of these tests. Results showed a concordance among the eight laboratories of 99.4%-100% for both arrays. The reproducibility of the tests was high with a relative standard deviation ranging from 1.1% to 7.1%. Considering the high level of agreement achieved between the laboratories these PCR arrays constitute robust and suitable tools for rapid detection of C. botulinum types C, D and mosaic types C-D and D-C. These are the first tests for C. botulinum C and D that have been evaluated in a European multicenter collaborative trial.
Anaerobe, 2013, Vol 22, p. 31-37
C. botulinum C and D; Animal botulism; Evaluation trial; GeneDisc array