90 likes | 178 Views
Summary of ACW work in Q-Detect. Andreas Bühlmann JF Pothier, THM Smits, JE Frey, B Duffy (ACW, Switzerland). Final Meeting Ljubljana – 18.02.2013. www.qdetect.org. XAP LAMP assay. Xanthomonas arboricola pv. pruni Bacterial spot of stone fruit ( Prunus ). Assay designed Validated
E N D
Summary of ACW work in Q-Detect Andreas Bühlmann JF Pothier, THM Smits, JE Frey, B Duffy (ACW, Switzerland) Final Meeting Ljubljana – 18.02.2013 www.qdetect.org
XAP LAMP assay Xanthomonas arboricola pv. pruni Bacterial spot of stone fruit (Prunus) • Assay designed • Validated • Ring tested • Paper published • EPPO validation Final Meeting Ljubljana – 18.02.2013
Ea LAMP assay Erwinia amylovora Fire blight of pome fruit (Spiraeoideae) • Assay designed • Validated • Field tested (only CH) • Paper published • EPPO validation • 96 full genomes confirm presence of diagnostic target Final Meeting Ljubljana – 18.02.2013
Further assays for plant pathogenic bacteria • Comparative genomics, primer design, validation with partners on… • Pantoea ananatis (ready for validation) • Pseudomonas syringae pv. actinida (genome sequenced, assay ready) • Xanthomonas oryzae pv. oryzae (Valerie Vérdier, F/USA) • Xanthomonas oryzae pv. oryzicola Final Meeting Ljubljana – 18.02.2013
Insect LAMP assays developed • Mostly on COI • Thrips palmi • Bemisia tabaci • Scirtothrips dorsalis • Bactrocera dorsalis group Final Meeting Ljubljana – 18.02.2013
Multiplexing LAMP • Swiss quarantine inspection service is considering Genies… • Would need: • Thrips palmi • Bactrocera spp. • Bemisisa tabaci () • Liriomyza trifolii/sativae (>80% of samples) • Multiplexable by melt curve: • Single tube assays make • inspectors lifes easier • Works with single insects Ea 86.0°C Bt 83.3°C Tp 81.8°C However - Multicolor still not done Final Meeting Ljubljana – 18.02.2013
Airport diagnostics CH • LAMP canimproveimportdiagnostics • Bemisiatabaciassayoptimized Final Meeting Ljubljana – 18.02.2013
SNP typing with LAMP • LAMP is not specific enough to detect SNPs • Same as TaqMan • LNA does not increase specificity but Speed. Final Meeting Ljubljana – 18.02.2013