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18-Month Meeting Padova (I) – 15-18.11.2011

Genomics-informed LAMP-PCR design for detection of Xanthomonas arboricola pv. pruni and Erwinia amylovora. Andreas Bühlmann JF Pothier, THM Smits, JE Frey, B Duffy (ACW, Switzerland). 18-Month Meeting Padova (I) – 15-18.11.2011. www.qdetect.org. Target Pathogens – EPPO A2 list bacteria.

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18-Month Meeting Padova (I) – 15-18.11.2011

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  1. Genomics-informed LAMP-PCR design for detection of Xanthomonas arboricola pv. pruni and Erwinia amylovora Andreas Bühlmann JF Pothier, THM Smits, JE Frey, B Duffy (ACW, Switzerland) 18-Month Meeting Padova (I) – 15-18.11.2011 www.qdetect.org

  2. Target Pathogens – EPPO A2 list bacteria Erwinia amylovora Fire blight of pome fruit (Spiraeoideae) Xanthomonas arboricola pv. pruni Bacterial spot of stone fruit (Prunus) Detection either by classical methods (slow), PCR, qPCR (expensive, limitations in field application), immunoassay (not very sensitive) Solution - LAMP Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

  3. Comparative genomics to identify targets Additional genomes sequenced/ added to analysis 4 Strains Xaj/Xaf 8 Strains Ea 7 Strains Pan/Ep Smits et. al. 2010 Smits et. al. 2010 Pothier et. al. unpublished Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

  4. Validation of targets XAP Specificity Sensitivy (Pure culture, inoculated plant samples) Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

  5. Validation of targets XAP Field samples In progress with CITA Zaragoza, IVIA Valencia Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

  6. Validation of targets Ea Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

  7. Validation of targets Ea Specificity Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

  8. Validation of targets Ea Sensitivity Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

  9. Validation of targets Ea Greenhouse/ field samples • 75 Samples of different apple varieties inoculated with Ea • 115 Samples naturally infected field samples from Switzerland • Samples from SI (in progress) Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

  10. On-site implementation of LAMP-PCR Integrating above methods with Q-Detect technology Equipment and on-site validation LAMP on LFD/ Genie Vs. Mobilab (classical portable PCR for Ea, XAP) Vs. Agristrip (Ea) Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

  11. On-site implementation of LAMP-PCR Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

  12. Off-topic LAMP Report on Swiss television on GenieII/LAMP on Thrips palmi developed at ACW Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

  13. Ring test for Xap LAMP in progress with CH, E and NL…. Ea LAMP to be tested in SI…. Collaboration on both LAMP tests with AUS (12.2011)…. Outlook Andreas Bühlmann – 15-18.11.2011 Q-Detect 18-Month Meeting, Padova I

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