A duplex quantitative real-time PCR assay for the detection and quantification of Xanthomonas phaseoli pv. dieffenbachiae from diseased and latently infected anthurium tissue
Anthurium bacterial blight caused by Xanthomonas phaseoli pv. dieffenbachiae (formerly Xanthomonas axonopodis pv. dieffenbachiae) is the major phytosanitary threat in many anthurium growing areas worldwide. Reliable and sensitive diagnostic tools are required for surveillance and certification programs. A duplex real–time quantitative PCR assay was developed for the detection and quantification of X. phaseoli pv. dieffenbachiae from anthurium tissue. This PCR assay targeted a X. phaseoli pv. dieffenbachiae–specific gene encoding an ABC transporter and an internal control encoding for chalcone synthase in Anthurium andreanum. A cycle threshold (Ct), using a receiver-operating characteristic approach (ROC), was implemented to ensure that the declaration of a positive sample was reliable. The duplex real–time assay displayed very high performance with regards to analytical specificity (100% inclusivity, 98.9% exclusivity), analytical sensitivity (LOD95% = 894 bacteria/ml corresponding to 18 bacteria per reaction) and repeatability. We demonstrated the pertinence of this real–time quantitative PCR assay for detecting X. phaseoli pv. dieffenbachiae from diseased leaf tissue (collected from outbreaks on anthurium) and from asymptomatic, latently infected anthurium plants. This assay could be useful for surveillance, as well as for indexing propagative plant material for the presence of X. phaseoli pv. dieffenbachiae.
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| Subjects: | H20 - Maladies des plantes, Anthurium, Xanthomonas campestris phaseoli, PCR, maladie bactérienne, diagnostic, infection latente, http://aims.fao.org/aos/agrovoc/c_487, http://aims.fao.org/aos/agrovoc/c_36124, http://aims.fao.org/aos/agrovoc/c_34079, http://aims.fao.org/aos/agrovoc/c_770, http://aims.fao.org/aos/agrovoc/c_2238, http://aims.fao.org/aos/agrovoc/c_16440, |
| Online Access: | http://agritrop.cirad.fr/592353/ http://agritrop.cirad.fr/592353/1/MIMET_jouen.pdf |
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dig-cirad-fr-5923532024-12-18T20:42:48Z http://agritrop.cirad.fr/592353/ http://agritrop.cirad.fr/592353/ A duplex quantitative real-time PCR assay for the detection and quantification of Xanthomonas phaseoli pv. dieffenbachiae from diseased and latently infected anthurium tissue. Jouen Emmanuel, Chiroleu Frédéric, Maillot-Lebon Véronique, Chabirand Aude, Mérion Sabine, Boyer Claudine, Pruvost Olivier, Robène Isabelle. 2019. Journal of Microbiological Methods, 161 : 74-83.https://doi.org/10.1016/j.mimet.2019.03.003 <https://doi.org/10.1016/j.mimet.2019.03.003> A duplex quantitative real-time PCR assay for the detection and quantification of Xanthomonas phaseoli pv. dieffenbachiae from diseased and latently infected anthurium tissue Jouen, Emmanuel Chiroleu, Frédéric Maillot-Lebon, Véronique Chabirand, Aude Mérion, Sabine Boyer, Claudine Pruvost, Olivier Robène, Isabelle eng 2019 Elsevier Journal of Microbiological Methods H20 - Maladies des plantes Anthurium Xanthomonas campestris phaseoli PCR maladie bactérienne diagnostic infection latente http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_770 http://aims.fao.org/aos/agrovoc/c_2238 http://aims.fao.org/aos/agrovoc/c_16440 Anthurium bacterial blight caused by Xanthomonas phaseoli pv. dieffenbachiae (formerly Xanthomonas axonopodis pv. dieffenbachiae) is the major phytosanitary threat in many anthurium growing areas worldwide. Reliable and sensitive diagnostic tools are required for surveillance and certification programs. A duplex real–time quantitative PCR assay was developed for the detection and quantification of X. phaseoli pv. dieffenbachiae from anthurium tissue. This PCR assay targeted a X. phaseoli pv. dieffenbachiae–specific gene encoding an ABC transporter and an internal control encoding for chalcone synthase in Anthurium andreanum. A cycle threshold (Ct), using a receiver-operating characteristic approach (ROC), was implemented to ensure that the declaration of a positive sample was reliable. The duplex real–time assay displayed very high performance with regards to analytical specificity (100% inclusivity, 98.9% exclusivity), analytical sensitivity (LOD95% = 894 bacteria/ml corresponding to 18 bacteria per reaction) and repeatability. We demonstrated the pertinence of this real–time quantitative PCR assay for detecting X. phaseoli pv. dieffenbachiae from diseased leaf tissue (collected from outbreaks on anthurium) and from asymptomatic, latently infected anthurium plants. This assay could be useful for surveillance, as well as for indexing propagative plant material for the presence of X. phaseoli pv. dieffenbachiae. article info:eu-repo/semantics/article Journal Article info:eu-repo/semantics/publishedVersion http://agritrop.cirad.fr/592353/1/MIMET_jouen.pdf text Cirad license info:eu-repo/semantics/restrictedAccess https://agritrop.cirad.fr/mention_legale.html https://doi.org/10.1016/j.mimet.2019.03.003 10.1016/j.mimet.2019.03.003 info:eu-repo/semantics/altIdentifier/doi/10.1016/j.mimet.2019.03.003 info:eu-repo/semantics/altIdentifier/purl/https://doi.org/10.1016/j.mimet.2019.03.003 |
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H20 - Maladies des plantes Anthurium Xanthomonas campestris phaseoli PCR maladie bactérienne diagnostic infection latente http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_770 http://aims.fao.org/aos/agrovoc/c_2238 http://aims.fao.org/aos/agrovoc/c_16440 H20 - Maladies des plantes Anthurium Xanthomonas campestris phaseoli PCR maladie bactérienne diagnostic infection latente http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_770 http://aims.fao.org/aos/agrovoc/c_2238 http://aims.fao.org/aos/agrovoc/c_16440 |
| spellingShingle |
H20 - Maladies des plantes Anthurium Xanthomonas campestris phaseoli PCR maladie bactérienne diagnostic infection latente http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_770 http://aims.fao.org/aos/agrovoc/c_2238 http://aims.fao.org/aos/agrovoc/c_16440 H20 - Maladies des plantes Anthurium Xanthomonas campestris phaseoli PCR maladie bactérienne diagnostic infection latente http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_770 http://aims.fao.org/aos/agrovoc/c_2238 http://aims.fao.org/aos/agrovoc/c_16440 Jouen, Emmanuel Chiroleu, Frédéric Maillot-Lebon, Véronique Chabirand, Aude Mérion, Sabine Boyer, Claudine Pruvost, Olivier Robène, Isabelle A duplex quantitative real-time PCR assay for the detection and quantification of Xanthomonas phaseoli pv. dieffenbachiae from diseased and latently infected anthurium tissue |
| description |
Anthurium bacterial blight caused by Xanthomonas phaseoli pv. dieffenbachiae (formerly Xanthomonas axonopodis pv. dieffenbachiae) is the major phytosanitary threat in many anthurium growing areas worldwide. Reliable and sensitive diagnostic tools are required for surveillance and certification programs. A duplex real–time quantitative PCR assay was developed for the detection and quantification of X. phaseoli pv. dieffenbachiae from anthurium tissue. This PCR assay targeted a X. phaseoli pv. dieffenbachiae–specific gene encoding an ABC transporter and an internal control encoding for chalcone synthase in Anthurium andreanum. A cycle threshold (Ct), using a receiver-operating characteristic approach (ROC), was implemented to ensure that the declaration of a positive sample was reliable. The duplex real–time assay displayed very high performance with regards to analytical specificity (100% inclusivity, 98.9% exclusivity), analytical sensitivity (LOD95% = 894 bacteria/ml corresponding to 18 bacteria per reaction) and repeatability. We demonstrated the pertinence of this real–time quantitative PCR assay for detecting X. phaseoli pv. dieffenbachiae from diseased leaf tissue (collected from outbreaks on anthurium) and from asymptomatic, latently infected anthurium plants. This assay could be useful for surveillance, as well as for indexing propagative plant material for the presence of X. phaseoli pv. dieffenbachiae. |
| format |
article |
| topic_facet |
H20 - Maladies des plantes Anthurium Xanthomonas campestris phaseoli PCR maladie bactérienne diagnostic infection latente http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_770 http://aims.fao.org/aos/agrovoc/c_2238 http://aims.fao.org/aos/agrovoc/c_16440 |
| author |
Jouen, Emmanuel Chiroleu, Frédéric Maillot-Lebon, Véronique Chabirand, Aude Mérion, Sabine Boyer, Claudine Pruvost, Olivier Robène, Isabelle |
| author_facet |
Jouen, Emmanuel Chiroleu, Frédéric Maillot-Lebon, Véronique Chabirand, Aude Mérion, Sabine Boyer, Claudine Pruvost, Olivier Robène, Isabelle |
| author_sort |
Jouen, Emmanuel |
| title |
A duplex quantitative real-time PCR assay for the detection and quantification of Xanthomonas phaseoli pv. dieffenbachiae from diseased and latently infected anthurium tissue |
| title_short |
A duplex quantitative real-time PCR assay for the detection and quantification of Xanthomonas phaseoli pv. dieffenbachiae from diseased and latently infected anthurium tissue |
| title_full |
A duplex quantitative real-time PCR assay for the detection and quantification of Xanthomonas phaseoli pv. dieffenbachiae from diseased and latently infected anthurium tissue |
| title_fullStr |
A duplex quantitative real-time PCR assay for the detection and quantification of Xanthomonas phaseoli pv. dieffenbachiae from diseased and latently infected anthurium tissue |
| title_full_unstemmed |
A duplex quantitative real-time PCR assay for the detection and quantification of Xanthomonas phaseoli pv. dieffenbachiae from diseased and latently infected anthurium tissue |
| title_sort |
duplex quantitative real-time pcr assay for the detection and quantification of xanthomonas phaseoli pv. dieffenbachiae from diseased and latently infected anthurium tissue |
| publisher |
Elsevier |
| url |
http://agritrop.cirad.fr/592353/ http://agritrop.cirad.fr/592353/1/MIMET_jouen.pdf |
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