Nested PCR Xad - 24 tests / Nested PCR Xad - 96 tests

Detection: Xanthomonas axonopodis pv. dieffenbachiae on anthurium.

Matrices: Bacterial culture or plant extracts, symptomatic and asymptomatic leaf.

Other information: kit developed and optimized by Qualiplante according to Robène-Soustrade et al. (2006). Under CIRAD license.

The PCR primers used in this Nested End-Point PCR method were selected from the SCAR sequence of Xad strain LMG695 and complemented some bases of the SCAR sequence.

This method is referred in the Appendix 4 of the PM7/23 (2) Xanthomonas axonopodis pv. dieffenbachiae, European and Mediterranean Plant Protection Organization Bulletin (2009) 39, 393-402.

The detection threshold obtained with pure cultures or plant extracts (103 CFU ml-1) allowed detection of the pathogen from symptomless contaminated plant. Internal validation data of the method are available on the ANSES/LSV/MA 028 – Version 3, September 2019 realized in 2008 by ANSES. The performance characteristics obtained are:

  • Sensitivity: 100% tested with 50 target bacterial strains in duplicate, with geographic and genetic diversity, diversity of host plants and years of isolation.
  • Specificity: 96% tested with 53 bacterial strains or non-target samples in duplicate.
  • Accuracy: 98% tested with 50 target bacterial strains in duplicate and 53 bacterial strains or non-target samples in duplicate.
  • Limit of detection: [5.103 – 5.104] mL-1.
  • Repeatability: 95% tested with 6 levels of contamination 0 – 102 – 103 – 104 – 105 -106 mL-1 x 4 different target strains representative of the diversity.
  • Reproducibility: 93% tested during an interlaboratory test comprising 15 different laboratories covering 4 different samples (8 replicates per sample and by laboratory) made up of ground healthy anthurium doped or not with targets or non-targets at different levels of contamination.

Kit referred in ANSES/LSV/MA 028.

Included reagents: Direct Master Mix, Nested Master Mix, Negative Control, Positive Control.


Targeted pathogen:

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