Folie 1

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Standardization of Automated Sample Preparation
for Viral Diagnostics
Dirk Heckel, PhD Diagnostic Sample Preparation
and Stabilization QIAGEN GmbH
2
Standardizing sample preparation in 2004

• QIAamp DSP Virus kit CE certified in April 2004
• BioRobot M48 for medium throghput in May 2004
• BioRobot MDx CE certification in Q3/04
• BioRobot EZ1 for low throughput in Q3/04

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Benefits of Standardization by Automation

• Automation of generic sample preparation methods
  can lead to
• Securing of result integrity
• Reduction of user errors
• Increased process control
• Comprehensive process documentation
• Medium to High throughput
• High level of user convenience

4
QIAGENs solution for required functionality (1)

• Automatic barcode reading of sample ID
• Tracking of sample ID from sample input area to
  result documentation
• Software guidance for system setup
• Automatic verification of complete and correct
  system setup (Load Check)
• Usage of filter tips
• 8-Channel Pipetting
• Proprietary design of robotic vacuum chamber
  (RoboVac)

• Sample ID tracking
• Secure system setup
• Cross contamination free processing

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QIAGENs solution for required functionality (2)

• Clot Check
• Positive sample dispense verification
• Liquid detection
• Constant verification of key process parameters
  (e.g. temperature/pressure)
• Load check
• Robotic arm with lab hand and 8 Channel dispenser
  head
• RoboVac
• Heating/Cooling device
• Result file including information such assample
  ID, sample validity, user, instrument, time
  stamp, maintenance status etc.

• High Level of Process control
• Walk away automation
• Result documentation

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Standardization Verification / Validation

• CE-Marking requires performance evaluation of the
  IvD system

System Verification Validation
Process Verification / HW/SW integration
verification
Chemistry Verification
Hardware Module Verification
Software Unit Level and Integration Level
Verification
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System Verification Validation
System Verification Validation

• Limit of detection
• Linearity and linear range
• Accuracy
• Precision, intermediate precision,
  reproducibility
• Robustness, whole system failure rate
• Cross contamination
• Clinical specificity

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Limit of detection QIAamp DSP 96 Virus MDx
Protocol
LOD HBV 26 IU/ml ? 144 c/ml
LOD HIV 123 IU/ml ? 70 c/ml
LOD HCV 28 IU/ml
all all all
Input Titer IU/ml all all all
Input Titer IU/ml n hits
800 36 36 100,00
600 36 36 100,00
400 36 36 100,00
200 36 36 100,00
100 36 35 97,22
50 36 23 63,89
10 36 6 16,67
NC 18 0 0,00
pos. control 18 18 100,00
95 probit value 95 probit value 123.1 123.1
confidence interval confidence interval 87.8-205.2 87.8-205.2
all all all
Input TiterIU/ml all all all
Input TiterIU/ml n hits
50 83 83 100,00
35 84 84 100,00
20 84 79 94,05
10 84 67 79,76
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   95 Probit value     25.70
confidence interval confidence interval confidence interval 19.37-37.18
all all all
Input Titer IU/ml all all all
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
10 36 35 97,22
5 36 23 63,89
0 36 6 16,67
95 probit value 95 probit value 28.2 28.2
confidence interval confidence interval 16,1-195.21 16,1-195.21
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Linear range (24 replicates at 7 titer levels)

• COBAS AMPLICOR HBV MONITOR

COBAS TaqMan HCV
LCx HIV RNA Quantitative Assay
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Intermediate Precision HIV cont.QIAamp DSP 96
Virus MDx Protocol
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Robustness

• Comparison of different primary tubes
• 4 Replicates of 13 individual donors
• 3 tube types per donor
• Spiked w/ 1105 IU/ml

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Standardizing sample preparation in 2004
BioRobot MDx DSP system

• QIAamp DSP Virus kit CE certified in April 2004
• BioRobot M48 for medium throghput in May 2004
• BioRobot MDx CE certification in Q4/04
• BioRobot EZ1 for low throughput in Q3/04

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BioRobot MDx DSP system
?

• BioRobot EZ1 CE certification in Q1 2006
• BioRobot EZ1 CE certification of Virus Kit in Q2
  2006