O28 Riedel

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2008 Phytophthora Workshop in Italy

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O28 Riedel

  1. 1. Nanobiotechnology for Phytophthora diagnosis M. Riedel, S. Julich, L. Dressler, R. Möller, S. Wagner, A. Breitenstein, T. Henkel and S. Werres which Phytophthora? Riedel et al. : Nanobiotechnology 3rd International Phytophthora, Pythium and related genera workshop 26.08.2008
  2. 2. aim of the project Development of a detection system - species specific - sensitive - robust - easy to handle - applicable in the field - results within short time chip-based diagnostic system Riedel et al. : Nanobiotechnology 3rd International Phytophthora, Pythium and related genera workshop 26.08.2008
  3. 3. functional principle of the chip system combined microchip for chip based PCR amplification (PCR chip) and DNA detection (DNA chip) via hybri- dization scheme of micro- systemic implemen- PCR chip DNA chip tation of automa- tized chip systems combined with an integrated micro- fluidic system for anlysis of nucleic acids buffer PCR complete control elements for waste pump wash solutions mix chip-system temperature control and enzymes electric detection Riedel et al. : Nanobiotechnology 3rd International Phytophthora, Pythium and related genera workshop 26.08.2008
  4. 4. PCR chip module on chip amplification und labeling of DNA by PCR single PCR chip bondpad temperature sensor detection window circular fluid anchor heater results: amplification of P. software PDA with control and analysis ramorum ITS2 fragment identical with conventional for chip-based PCR amplification thermocycler and PCR-chip Riedel et al. : Nanobiotechnology 3rd International Phytophthora, Pythium and related genera workshop 26.08.2008
  5. 5. DNA chip module detection of Phytophthora DNA fragments on chip Phytophthora specific DNA of closely first results: hybridization probes related Phytophthora taxa spotted at elektrode gap (10 µm) unspecific hybridization: P. lateralis : P. ramorum example for species specific probes :: example for species specific probes P.ramorum P.ramorum GGAAACTTGT GTTGGCGGCT TCGGCTGCTG GGAAACTTGT GTTGGCGGCT TCGGCTGCTG P.lateralis GGAAATTTGT GTTGGCAGCT TCGGCTGTTG P.lateralis GGAAATTTGT GTTGGCAGCT TCGGCTGTTG specific hybridization: detection of hybrisization results by silver enhancement - electrical P. ramorum P. ramorum : or optical analysis possible combined gold chip with 42 detec- tion sites and microfluidic Riedel et al. : Nanobiotechnology 3rd International Phytophthora, Pythium and related genera workshop 26.08.2008
  6. 6. Acknowledgements We would like to thank the Bundesanstalt für Ernährung und Landwirtschaft (BLE) for funding our project and all partners at JKI, IPHT und BECIT ! Riedel et al. : Nanobiotechnology 3rd International Phytophthora, Pythium and related genera workshop 26.08.2008

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