Development of advanced and innovative diagnostic assays for the detection of Xylella fastidiosa in plants and insect-vectors
Résumé
Although DNA- and protein-based methods have revolutionized Xylella fastidiosa diagnosis, in some cases they are not very reliable due, on one hand, to the peculiarities of this pathogen, i.e. (i) irregular distribution inside the plant, (ii) xylem-limited localization making its extraction difficult, (iii) latency in some species that can affect accurate sampling; (iv) low concentration in some hosts; (v) genomic variance between subspecies; and, on the other hand, the poor performance of some serological and molecular techniques to sensitively and differentially detect and discriminate between subspecies and strains of Xylella fastidiosa.
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Accordingly, the aim of this study was to develop advanced serological (Nanobodies) and molecular (Real-time TaqMan PCR, LAMP and FTPS-LAMP) assays able to detect Xylella fastidiosa, especially the CoDiRO strain present in Apulia, in an easy, sensitive, accurate and discriminating manner. The hemagglutinin gene, found solely in the CoDiRO genome, together with two universal genes of Xylella fastidiosa, were used for the construction of protein- and DNA-based techniques, respectively. These three recombinant proteins were bacterially expressed, purified and quantified for future injections in rabbit and Llama for the production of polyclonal and monoclonal antibodies against Xylella fastidiosa. Real-time TaqMan PCR, LAMP and FTPS-LAMP assays here developed in singleplex and multiplex approaches were found highly sensitive, specific and discriminant to detect Xylella fastidiosa in infected plants and insect-vectors.
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Bibliothèque
Biblioteca CIHEAM Bari
Document à consulter sur place uniquement
Cote
ARCH M.Sc. 883 2019
Document à consulter sur place uniquement
Cote
M.Sc. 883 2019
Document à consulter sur place uniquement
Cote
M.Sc. 883 A 2019