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Automated RNA/DNA Extraction with TaqMan Technology

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Process and Develop Analytical Solutions for Efficient RNA and DNA Extraction Combined with TaqMan Technology. Validation according to GMPs. Achieve Detection Limits for HAV, HCV, HBV, HIV-1, and Parvovirus B19.

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Automated RNA/DNA Extraction with TaqMan Technology

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  1. Automated Extraction for RNA/DNA Combined with TaqMan Technology and Cobas AmpliscreenElisa Moretti Process and Analytical DevelopmentKEDRION S.p.ASoGAT XVIPaul Ehrlich Institut - Langen, Germany3rd July 2003

  2. Virus Kit (BIOROBOT 9604 QIAGEN) AMPLIFICATION & DETECTION HIV-1, HBV & HCV PVB19 & HAV KIT Cobas Ampliscreen (Cobas Amplicor) ROCHE In-house Protocol (ABI PRISM 7900HT) APPLERA NAT Development Single extraction for RNA & DNA

  3. Final Results Instruments validation according to GMPs HAV: Detection Limit: 33,68 UI/ml 95% C.I. 25,86 - 49,86 UI/ml HAV-RNA ISS 0201 Preliminary Results HCV 12 UI/ml = 100 % WHO IS 96/790 NIBSC HIV-1 100 UI/ml = 100% WHO IS 97/656 NIBSC HBV 6 UI/ml = 100 % WHO IS 97/746 NIBSC

  4. Parvovirus B19 • Detection Limit: • 133,35 UI/ml • 95% C.I. • 107,63 - 185,77 • Linearity: • 105-103 UI/ml • Cross-contamination: • Problems with high • PVB19 titre • WHO IS 99/800 NIBSC • Alternative extraction • QIAamp MinElute Virus • Spin Kit or Ampliscreen • Amplification & Detection • ABIPrism 7900HT • Evaluation in progress • Validation: • Quantitative Method • (PA/PH/OMCL (03) 38 2R; June 2003)

  5. Acknowledgements • Laboratory of Immunology ISS • Giuliano Gentili • German Red Cross, Frankfurt • Willi Kurt Roth • Kedrion • Giampaolo Nardi

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