Finally the LFD strip was submerged into Ku-0059436 supplier the mixture, and the results were visualized after 5 minutes. Sensitivity of LAMP and real time PCR In order to estimate the sensitivity of the Las-LAMP assay, purified DNA from a Las infected plant was serially diluted and 1 μL aliquots of these dilutions were used as template for Las-LAMP and real time PCR. Las-LAMP reactions were performed as mentioned above, and real time PCR was carried out as described previously [3], in a Step One™ real time PCR system (Applied Biosystems®). Acknowledgements

We thank Dr. Keith Ireton for critical review of the manuscript. We are grateful to Dr. Nelson Arno Wulff of Fundecitrus, São Paulo, Brazil, for providing some of the DNA samples used in this study. Thanks to OPENCLIPART.ORG for providing community sourced images that were used to create illustrations in this manuscript. MRM, APC and AAV are Career Investigators of Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina. This work was supported by Agencia de Promoción Científica y Tecnológica of Argentina. Electronic supplementary

material Additional file 1: Figure S1: Pairwise alignment between CLIBASIA_05175 and related sequences from a BLASTn search. A. BLASTn pairwise alignment between CLIBASIA_05175 (green) and a related sequence from Candidatus Liberibacter solanacearum (black). Las-LAMP primer binding sites are highlighted in yellow and cyan. B. BLASTn pairwise alignment between https://www.selleckchem.com/products/Fedratinib-SAR302503-TG101348.html CLIBASIA_05175 (green) and a related sequence from Candidatus Liberibacter americanus (black). Las-LAMP primer binding sites are highlited in yellow and cyan. C. BLASTn pairwise alignment between CLIBASIA_05175 (green) isometheptene and a related sequence from Candidatus crescens (black). Las-LAMP primer binding sites are highlighted in yellow and in cyan. (PPTX 540 KB) Additional file 2: Figure S2: Evaluation of Candidatus Liberibacter americanus DNA by Las-LAMP. Purified DNA from plants infected with Candidatus Liberibacter asiaticus (Las) or Candidatus Liberibacter

americanus (Lam) were used as templates for the Las-LAMP amplification reaction. A. Amplification products analyzed by gel electrophoresis. B. Amplification products analyzed using a lateral flow dipstick. C-: negative control without template. M: 1 Kb plus DNA ladder (Invitrogen®), the size of the bands is, from bottom to top: 100 bp, 200 bp, 300 bp, 400 bp, 500 bp, 650 bp, 850 bp, 1000 bp, 1650 bp, 2000 bp and increments of 1000 bp up to 12000 bp. (PPTX 2 MB) Additional file 3: Figure S3: Pairwise alignment between CLIBASIA_05175 and the sequence of a Las-LAMP amplification product. A Las-LAMP amplification product band was Analyzed by sequencing. The sequence corresponding to the amplification product (red), from F2 to B2c, has been subjected to a pairwise alignment against CLIBASIA_05175 (green). (PPTX 134 KB) Additional file 4: Figure S4: Evaluation of different heating devices on Las-LAMP amplification.