w w w. q i a g e n. c o m dirk heckel, phd diagnostic sample preparation and stabilization qiagen...
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W W W . Q I A G E N . C O M
Dirk Heckel, PhDDirk Heckel, PhD
Diagnostic Sample Preparation and StabilizationDiagnostic Sample Preparation and Stabilization
QIAGEN GmbHQIAGEN GmbH
Standardization of Standardization of Automated Sample Automated Sample
Preparation for Preparation for Viral DiagnosticsViral Diagnostics
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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
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QIAGEN’s solution for required functionality (1)
Sample ID tracking
Secure system setup
Cross contamination free processing
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)
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QIAGEN’s solution for required functionality (2)
High Level of Process control
Walk away automation
Result documentation
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.
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Standardization = Verification / Validation
CE-Marking requires performance evaluation of the IvD system
BioRobot MDx DSP system
IvD ReagentsIvD
InstrumentApplication
process
IvD Hardware IvD SoftwareHardware Module VerificationSoftware Unit Level
and Integration Level Verification
Process Verification / HW/SW integration
verificationChemistry Verification
System Verification & Validation
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System Verification & Validation
Limit of detection
Linearity and linear range
Accuracy
Precision, intermediate precision, reproducibility
Robustness, whole system failure rate
Cross contamination
Clinical specificity
System Verification & Validation
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Limit of detection QIAamp DSP 96 Virus MDx Protocol
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 123.1
confidence interval 87.8-205.2
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
25.70
confidence interval 19.37-37.18
LOD HBV = 26 IU/ml
144 c/ml
LOD HIV = 123 IU/ml 70 c/ml
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 28.2
confidence interval 16,1-195.21
LOD HCV = 28 IU/ml
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Linear range (24 replicates at 7 titer levels)
y = 1.0577x - 0.5599
R2 = 0.9987
0
1
2
3
4
5
0 1 2 3 4 5Input (log IU/ml)
Resu
lt (lo
g IU
/ml)
COBAS AMPLICOR HBV MONITOR
y = 0.9054x + 0.3011
R2 = 0.9982
1
2
3
4
5
6
7
1 2 3 4 5 6 7Input (log IU/ml)
Resu
lt (lo
g IU
/ml)
y = 0.8867x + 0.3878
R2 = 0.9972
2
3
4
5
6
7
8
2 3 4 5 6 7 8
Input (log IU/ml)
Res
ult (lo
g IU
/ml)
COBAS TaqMan HCV
LCx HIV RNA Quantitative Assay
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0,00
1,00
2,00
3,00
4,00
5,00
6,00
7,00
8,00
log c
/ml
Dilution 1 Dilution 2 Dilution 3 Dilution 4 Dilution 5 Dilution 6 Dilution 7
QIAamp MDx DSP 96 Virus MDx protocol - Intermediate Presision
Day1, System1
Day1, System2
Day2, System1
Day2, System2
Day2, System3
Day3, System2
Day3, System3
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/ 1*10^5 IU/ml
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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 Q4/04 BioRobot EZ1: for low throughput in Q3/04 BioRobot EZ1: CE certification in Q1 2006 BioRobot EZ1: CE certification of Virus Kit in Q2 2006
2005
BioRobot MDx DSP system
BioRobot MDx DSP system