Recommendations from Euphresco projects
The following research project has recently been carried out in the framework of Euphresco (network for phytosanitary research coordination and funding - hosted by EPPO). A report presenting the main objectives and results of this project, as well as recommendations made can be viewed on the Internet.
Interlaboratory test performance studies for identification of Ralstonia solanacearum and molecular confirmation of its virulence
The EU directive 2006/63/EC describes a detailed protocol for the official testing of Ralstonia solanacearum that is internationally recognized and has been implemented in many diagnostic laboratories across Europe and beyond. In this protocol, the confirmation of the identity of the bacterium is performed by a laborious, time-consuming and expensive pathogenicity test. The aim of this project was to develop and evaluate molecular diagnostic methods for the detection and identification of Ralstonia solanacearum, as well as for the verification of its virulence, that would be faster, more specific, and robust.
The consortium was not successful in the identification of virulence genes to be considered for the development of the diagnostic test. A comparison of a real-time LAMP test, a real-time PCR test, a conventional PCR test and an Immunofluorescence Antibody Staining (IFAS) test was undertaken. Real-time LAMP was shown to be the most sensitive diagnostic method. The test also offers numerous advantages: it is rapid, cost-efficient, user-friendly and less influenced by inhibitors derived from culture medium, plant tissues and soil compared to PCR. In addition, it requires less stringent DNA extraction procedures than those for conventional PCR and real-time PCR. It was concluded that the LAMP test could be a valuable tool for the testing of Ralstonia solanacearum and should be recommended in international standards. More work is needed to improve the DNA extraction methods.
Duration of the project: 2018-05-01 to 2020-10-29.
Authors: Tjou-Tam-Sin, Napoleon; Vogelaar, Martijn; Li, XIang; Čermák, Vaclav; Fornefeld, Eva; van der Wolf, Jean Martin; Valentini, Franco; Fraser, Karen; Cara, Magdalena; Yildiz, Nilufer; Yuzbasioglu, Eda; Karahan, Aynur; Ustun, Nursen; Kreuze, Jan.
Link: https://zenodo.org/record/4153357#.X5qfk4hKiUk
Sources
Euphresco (2020-11). https://www.euphresco.net/projects/ |