Rapid loop-mediated isothermal amplification assays for grapevine yellows phytoplasmas on crude leaf-vein homogenate has the same performance as qPCR |
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| Authors: | Polona Kogov?ek Nata?a Mehle Anja Pugelj Tja?a Jakomin Hans-Josef Schroers Maja Ravnikar Marina Dermastia |
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| Institution: | 1.Department of Biotechnology and Systems Biology,National Institute of Biology,Ljubljana,Slovenia;2.Agricultural Institute of Slovenia,Ljubljana,Slovenia;3.University of Nova gorica,Nova Gorica,Slovenia |
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| Abstract: | A fluorescence-based real-time loop-mediated isothermal amplification (LAMP) assay for ‘Candidatus Phytoplasama solani’ (Bois noir phytoplasma; BNp) detection was developed and optimised for rapid laboratory and on-site BNp detection. This assay is highly specific, rapid and as sensitive as qPCR. It was validated according to European and Mediterranean Plant Protection Organisation recommendations. In addition, 286 grapevine leaf samples from the 2015 growing season were tested with this new real-time LAMP assay and an assay previously developed for detection of Flavescence dorée phytoplasma (FDp). These LAMP assays for detection of both BNp and FDp used without any DNA extraction step, which is a required step for qPCR analysis, were comparably effective to qPCR, and positive results were obtained in less than 35 min. |
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