2. DRK-Blutspendedienst West  nPrecipitation steps (PEG), centrifugation and careful resuspension of the invisible pelleted viral particles precede efficient extraction of viral RNA and DNA from minipools (n  96,  9.6 ml). Loss of nucleic acid during this time consuming steps is another critical point. nWe looked for an automated RNA/DNA extraction method, that could start right from the minipool sample (n = 96, 9.6 ml). Release of labile blood components calls for short assay time. Zentrallabor Hagen SoGAT XVII, Paris, 27/05/04 Magnetic bead technology in viral RNA/DNA extraction from plasma minipools L. Pichl and V. Schottstedt German Red Cross Blood Transfusion Centre West, Central Laboratory, Hagen

3. DRK-Blutspendedienst West  Magnetic Separation Module I SoGAT XVII, Paris, 27/05/04 Zentrallabor Hagen Rod Head Electromagnet Tube track

4. DRK-Blutspendedienst West  Features nPolyethanol coated magnetic beads (1 µm Ø, hydrophilic) n12 Rod Head magnetizable n12 samples, up to 10 ml vol. in 50 ml tubes nOne specific disposable tip per sample per complete separation nEluate volume: 100 µl nProcessing time: 1h 8 min SoGAT XVII, Paris, 27/05/04 Zentrallabor Hagen

5. DRK-Blutspendedienst West  Materials nChemagen Magnetic Separation Module I nChemagic Viral 10k Kit Special (www.chemagen.com) nLightCycler II (Roche Diagnostics), ABI SDS 7700 nRealArt™ ParvoB19 LC PCR Kit RealArt™ HAV LC RT PCR Kit RealArt™ HBV TM PCR Kit (www.artus-biotech.com) Nucleic acid extraction Amplification and detection SoGAT XVII, Paris, 27/05/04 Zentrallabor Hagen

6. DRK-Blutspendedienst West  Workflow SoGAT XVII, Paris, 27/05/04 Zentrallabor Hagen

7. DRK-Blutspendedienst West  Sensitivity Magnetic Separation Module I + LightCycler II /ABI SDS 7700 Minipools of n = 93, EDTA-Plasma, triple-spike of diluted Ref. Mat., 100 µl each SoGAT XVII, Paris, 27/05/04 Zentrallabor Hagen

8. DRK-Blutspendedienst West  Robustness n102 Pools have been analysed for B19, HBV and HAV so far. None of them tested positive. nCross contamination study was performed with two 3 x 4 matrices of alternating negative and B19 spiked pools. Referring to a dose of 10EE 8 IU/ml per single donation all of the none spiked pool were negative in B19 LC-PCR. nTracing back reactive results from pools to single donation via ”chessboard“ testing was performed for HAV, HBV and B19. SoGAT XVII, Paris, 27/05/04 Zentrallabor Hagen

9. DRK-Blutspendedienst West  Outlook nCombination of Chemagen´s magnetic bead tech- nology with real time PCR without pre-extraction manipulation of the minipool (n = 96) reveals acceptable sensitivity and robustness for PAV B19, HAV and HBV; nImplementation of a robotic sample processor with the magnetic separation module for sample identification, aliquoting reagents and sample lysis; nIncreasing throughput by optimizing time management. SoGAT XVII, Paris, 27/05/04 Zentrallabor Hagen

10. DRK-Blutspendedienst West  Thanks to: Thorsten Hageböck Andrea Matulina Yvonne Schmidt Zentrallabor Hagen SoGAT XVII, Paris, 27/05/04