These authors contributed equally to this work.
Comparative and collaborative studies for the validation of a nested PCR for the detection of Xanthomonas axonopodis pv. dieffenbachiae from Anthurium samples
Article first published online: 17 JUN 2013
© 2013 British Society for Plant Pathology
Volume 63, Issue 1, pages 20–30, February 2014
How to Cite
Chabirand, A., Jouen, E., Pruvost, O., Chiroleu, F., Hostachy, B., Bergsma-Vlami, M., Bianchi, G., Cozzolino, L., Elphinstone, J., Holeva, M., Manole, F., Martini, P., Matoušková, H., Minatchy, J., de Beeck, G. O., Poliakoff, F., Sigillo, L., Siverio, F., Van Vaerenbergh, J., Laurentie, M. and Robène-Soustrade, I. (2014), Comparative and collaborative studies for the validation of a nested PCR for the detection of Xanthomonas axonopodis pv. dieffenbachiae from Anthurium samples. Plant Pathology, 63: 20–30. doi: 10.1111/ppa.12083
- Issue published online: 7 JAN 2014
- Article first published online: 17 JUN 2013
- Manuscript Accepted: 22 APR 2013
- Anses Plant Health Laboratory
- Anthurium ;
- interlaboratory study;
- ISO 16140;
- PCR ;
Efficient control of Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of anthurium bacterial blight, requires sensitive and reliable diagnostic tools. The European standard EN ISO 16140:2003 has been followed to compare a nested PCR assay (N-PCR) to a reference method (isolation and serological identification of bacterial colonies) and to other alternative serological detection methods. The evaluation was performed in two steps: a comparative study and a collaborative study involving 15 European laboratories. Although inclusivity was maximal (100%) for all methods, a maximal exclusivity was obtained only with N-PCR followed by an enzymatic restriction digestion of the amplicons. Exclusivity indices of 90·6, 88·7 and 47·2% were found for indirect ELISA, immunofluorescence and double antibody sandwich ELISA, respectively. An exclusivity of 92·5% was obtained with the reference method, further increased to 100% if pathogenicity tests were performed as a supplemental assay. The best level of sensitivity (relative detection level) was obtained with the reference method followed by the N-PCR assay. The N-PCR performance in terms of relative accuracy, accordance and concordance was very similar to that of the reference method. Moreover, N-PCR had undeniable advantages compared to the reference method (less labour-intensive and less time-consuming). In addition, post-test probabilities of infection were calculated to select the most appropriate detection scheme related to the prevalence of the pathogen. The N-PCR assay has since been included in a revised version of the EPPO detection protocol.