Standardization of Automated Sample Preparation for Viral Diagnostics

Standardization of Automated Sample Preparation for Viral Diagnostics Dirk Heckel, Ph. D Diagnostic Sample Preparation and Stabilization QIAGEN Gmb. H W W W. Q I A G E N. C O M

Standardizing sample preparation in 2004 QIAamp DSP Virus kit: CE certified in April 2004 Bio. Robot M 48 for medium throghput in May 2004 Ø Bio. Robot MDx: CE certification in Q 3/04 Ø Bio. Robot EZ 1: for low throughput in Q 3/04 W W W. Q I A G E N. C O M

Benefits of Standardization by Automation of generic sample preparation methods can lead to: n Securing of result integrity n Reduction of user errors n Increased process control n Comprehensive process documentation n Medium to High throughput n High level of user convenience W W W. Q I A G E N. C O M

QIAGEN’s solution for required functionality (1) n Sample ID tracking n Automatic barcode reading of sample ID n Tracking of sample ID from sample input area to result documentation n Secure system setup n Software guidance for system setup n Automatic verification of complete and correct system setup (Load Check) n Cross contamination free processing n Usage of filter tips n 8 -Channel Pipetting n Proprietary design of robotic vacuum chamber (Robo. Vac) W W W. Q I A G E N. C O M

QIAGEN’s solution for required functionality (2) n High Level of Process control n Clot Check n Positive sample dispense verification n Liquid detection n Constant verification of key process parameters (e. g. temperature/pressure) n Walk away automation n Load check n Robotic arm with lab hand 8 Channel dispenser head n Robo. Vac n Heating/Cooling device n Result documentation n Result file including information such as sample ID, sample validity, user, instrument, time stamp, maintenance status etc. W W W. Q I A G E N. C O M

Standardization = Verification / Validation CE-Marking requires performance evaluation of the Iv. D system System Verification & Bio. Robot MDx DSP system Validation Iv. D Reagents Chemistry Verification Process Verification / Application Iv. D Instrument. HW/SW integration process verification Hardware Module. Iv. D Verification Software Unit Level Iv. D Software and Integration Level Verification W W W. Q I A G E N. C O M

System Verification & Validation § Limit of detection § Linearity and linear range § Accuracy § Precision, intermediate precision, reproducibility § Robustness, whole system failure rate § Cross contamination § Clinical specificity W W W. Q I A G E N. C O M

Limit of detection QIAamp DSP 96 Virus MDx Protocol LOD HBV = 26 IU/ml LOD HIV = 123 IU/ml 70 c/ml 144 c/ml all all % Input Titer IU/ml n hits % 83 100, 00 800 36 36 100, 00 84 84 100, 00 600 36 36 100, 00 20 84 79 94, 05 400 36 36 100, 00 10 84 67 79, 76 200 36 36 100, 00 100 36 35 97, 22 50 36 23 63, 89 10 36 6 NC 18 pos. control 18 Input Titer IU/ml n hits 50 83 35 5 84 50 59, 52 2 84 27 32, 14 0, 5 84 10 11, 90 0 41 0 0, 00 pos. control 42 42 100, 00 95% Probit value confidence interval LOD HCV = 28 IU/ml 25. 70 19. 37 -37. 18 Input Titer IU/ml n hits % 160 36 36 100, 00 80 36 36 100, 00 40 36 36 100, 00 20 36 36 100, 00 16, 67 10 36 35 97, 22 0 0, 00 5 36 23 63, 89 18 100, 00 0 36 6 16, 67 95% probit value 123. 1 95% probit value 28. 2 confidence interval 87. 8 -205. 2 confidence interval 16, 1 -195. 21 W W W. Q I A G E N. C O M

Linear range (24 replicates at 7 titer levels) COBAS AMPLICOR HBV MONITOR COBAS Taq. Man HCV LCx HIV RNA Quantitative Assay W W W. Q I A G E N. C O M

Intermediate Precision HIV cont. QIAamp DSP 96 Virus MDx Protocol W W W. Q I A G E N. C O M

Robustness § § Comparison of different primary tubes 4 Replicates of 13 individual donors 3 tube types per donor Spiked w/ 1*10^5 IU/ml W W W. Q I A G E N. C O M

Standardizing sample preparation in 2004 2005 Bio. Robot MDx DSP system QIAamp DSP Virus kit: CE certified in April 2004 Bio. Robot M 48 for medium throghput in May 2004 Ø Bio. Robot MDx: CE certification in Q 4/04 Bio. Robot MDx DSP system Ø Bio. Robot EZ 1: for low throughput in Q 3/04 Ø Bio. Robot EZ 1: CE certification in Q 1 2006 Ø Bio. Robot EZ 1: CE certification of Virus Kit in Q 2 2006 W W W. Q I A G E N. C O M