Novel tetraplex qPCR assays for simultaneous detection and identification of Xylella fastidiosa subspecies in plant tissues

Dupas Enora; Briand Martial; Jacques Marie-Agnès; Cesbron Sophie

solutions. The XF, XFM, XFMO, XFP, and XFFSL primers and probes allowed detection of Xf up to 278 10 pg.mL -1 (4 copies/reaction). XFF primers were slightly less sensitive with a threshold up to 100 279 pg.mL -1 (40 copies/reaction primers (XFMO) the LOD of tetraplex qPCR assays was usually 10 times higher than the LOD of the 289 simplex test on DNA (Table 4 and Supplemental data 2). In addition, it should be noted that the closer 290 the Ct value was to the detection limit, the higher the SEM was. In tetraplex qPCR assays set n°1, XF, 291 . CC-BY-NC-ND 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/699371 doi: bioRxiv preprint in planta identification of Xylella fastidiosa subspecies 8 XFM and XFP primers allowed a detection up to 100 pg.mL -1 . The XFFSL primers allowed the 292 detection of Xff up to 10 pg.mL -1 and of Xfmo up to 100 pg.mL -1 . The set n°2 allowed detection up to 293 100 pg.mL -1 using XFF and XFM primers and up to 10 pg.mL -1 with XFP primers. The XF primers 294 allowed the detection of Xff and Xfm up to 100 pg.mL -1 and of Xfp up to 10 pg.mL -1 . The set n°3, 295 allowed a detection up to 100 pg.mL -1 with XF, XFF and XFM primers and up to 10 pg.mL -1 with 296 XFMO primers. 297