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

2. 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
x 96 # samples

3. 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

4. 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

5. 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

6. 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

7. 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.

8. Thank You
for Your Attention!