Generic Automated Sample Preparation for Commercial NAT Assays

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Presentation transcript:

Generic Automated Sample Preparation for Commercial NAT Assays Markus Sprenger-Haussels, PhD Associate Director R&D QIAGEN GmbH, Hilden, Germany 14.06.2006 SoGAT XIX, Bern, Switzerland

Automation of Human Sample Preparation Chemistry + Application Expertise + Instrumentation BioRobot EZ1 Magnetic Beads BioRobot M48 Magnetic Beads BioRobot MDx Silica Membranes Protedyne BioCube Membranes / Beads 1 6 48 96 x 96 # samples

Parameters Influencing NAT Assays Preanalytical Assay/Detection Starting material is different liquid samples (plasma, serum, CSF, urine) solid samples (tissue, swabs, blood cards, stool) mechanical forces needed for disruption enzymatic forces required thermal lysis required chemical lysis sufficient rich in protein rich in polysaccharides rich in salts rich in everything rich in inhibitory metabolites interfering medical treatment Viruses are different single stranded double stranded RNA Viruses DNA Viruses linear genome circular genome degree of secondary structure enveloped or not lysis behavior low to high copy no. NA heat labile or not Different assays artus® Assays Roche Amplicor® Roche Cobas®TaqMan Abbott LCx® Abbott Taqman Bayer bDNA Bayer TMA Innogentics InnoLipa Homebrew many other in Generic Sample Preparation Need to be Neutralized Different controls “naked” RNA amored RNA plasmids linear DNA armored DNA virus like particles different length

Example: Magnetic Bead Technology Technology used for automation High number of optional modifications Size Weight Core material Surface material Surface structure Movement in liquids Physical change by protocol steps Differential binding of different nucleic acid types

New Automated Viral Sample Preparation Large Number of Assays covered Various Assays used during Development: artus Assays HBV*, EBV*, HSV*, HAV*, CMV* other artus Assays QuantiTect PCR Kits Homebrew Virus Assays Other Commerical Assays Roche Assays Abbott Assays Bayer Assays Others BioRobot EZ1 BioRobot M48 * as CE marked IVD only available in Europe

New Automated Viral Sample Preparation Link to Real-Time PCR Assays (1) EZ1 DSP Virus Kit with artus® HBV PCR Kits (CE)* 95 % probit values on different detection platforms 13 different dilution levels; 18 replicates per level LightCycler® 45.7 IU/ml TaqMan® 13.2 IU/ml Rotor-GeneTM 14.4 IU/ml * artus HBV PCR Kits only available in Europe as CE-IVDs; EZ1 DSP Virus Kit under CE-IVD development

New Automated Viral Sample Preparation Link to Real-Time PCR Assays (2) EZ1 DSP Virus Kit* with other NAT Assays: first verification LOD data with limited replicates *Under development for in-vitro-diagnostic use in Europe, not available in the U.S.A.

Thank You for Your Attention!