COBAS TaqMan 48 Sales Brochure

COBAS TaqMan 48 Product Presentation

Evolution of Roche Diagnostics Platforms (RMD)

COBAS AMPLIPREP& COBAS TaqMan 96/ TaqMan 48

Degree of Automation

COBAS TaqMan 48
COBAS AMPLIPREP Automated Sample Preparation AMPLILINK COBAS AMPLICOR MONITOR COBAS AMPLICOR

MWP

1992

1995

1996

2001

2003

COBAS TaqMan 48
First Real Time PCR Platform that Offers IVD & Utility Channel mode Programmable Benchtop analyzer 2 independent 24-well Thermal Cyclers (two tests simultaneously)

AmpliLink 3.0.1 Software Windows XP operating Sytems Run sizes of 6-48 samples

COBAS TaqMan with open thermalcycler

COBAS TaqMan Analyzers Common Features and Benefits

Feature
Kinetic PCR

Benefit
Broad dynamic range No separate detection Assay time reduction from 6h to 2-3 hours Increases Throughput Provides internal control
Eliminates contamination risk Allows multiplexing Will allow additional formats (Hybridization Probes) Easy handling Automation through AmpliPrep

Integrated Quantification Standard requires no external standard curves Closed Tube Format Four color channels (upgrade to six planned) Sample vessel: k-tube or kplate

COBAS TaqMan 48 Thermal Cyclers

TC lid

TC cover

K-tube, K-carrier, K-carrier holder

K-carrier Holder

K-carrier with barcode

K-tube

COBAS TaqMan 48
K-carrier Transporter

COBAS TaqMan 48

Motorized K-tube Capper

Post Launch improvements

K-Plate & K-caps in new K-carrier design (in development)

High Pure System Viral Nucleic Acid Kit Overview

COBAS TaqMan 48 Workflow with manual or automated sample prep

Manual Sample Prep

September 2003

Automated Sample Prep

July 2004

High Pure System Viral Nucleic Acid Kit

(redesigned - 12 samples format)
1 = Lysis Rack 2 = Filter Tube Rack 3 = Waste Container

4 = Elution Rack 5 = Cover Rack 6 = Grippers

One kit = 48 preps 4 plates with 12 Columns per plate; Lysis, Filter, Elution Rack

High Pure System Viral Nucleic Acid Kit/COBAS TaqMan TM HBV Test Performance

High Pure System / COBAS TaqMan HBV Linearity/Dynamic Range Study

Kit / COBAS TaqMan HBV Test Result Log10 (HBV DNA IU/mL) High Pure System Viral Nucleic Acid
9.00 8.00 7.00 6.00 5.00 4.00 3.00 2.00 y = 1.0124x - 0.2358 R2 = 0.9989 Error Bars = 1 Standard Deviation

1.00
0.00 -1.00 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00

Nominal Concentration Log10 (HBV DNA IU/ml)

HPS/CTM HBV linear range = 30 - 1.1E8 IU/ml

High Pure System / COBAS TaqMan HBV - Limit of Detection Study

EDTA-Plasma # Replicates # Positive Positivity Results Rate (%) 120 120 120 120 120 120 120 120 118 119 106 100 67 58 100 98 99 88 83 56 48 Serum # Positive Results 120 111 111 104 73 47 30

HBV Concentration (IU/ml) 12.7 8.5 5.9 3.4 17 0.8 0.4

# Replicates

Positivity (%) 100 95 97 87 64 39 25

120 117 114 120 114 120 120

HPS/CTM HBV can detect HBV DNA in EDTA-plasma and serum at concentrations as low as 5.9 IU/ml with a positivity rate greater than 95%

Roche HBV MONITOR Tests Comparison

Test/kit Batch size Sample type Sample volume Limit of detection Linear range Reportable units Genotype inclusivity Number of controls Quantitation standard AMPLICOR HBV MONITOR 48 16 serum & plasma 50 l 1000 copies/ml 1000 4E7 copies/ml copies/ml COBAS AMPLICOR HBV MONITOR 48 12 serum & plasma 100 l 300 copies/ml 300 200,000 copies/ml copies/ml (fixed conversion factor to convert results to IU/ml A-E; limited F&G recovery 3 (negative, low & high positives) Yes, QS incorporated into each sample prior extraction ~ 8 hours HPS/CTM HBV 48 12 serum & plasma 500 l 6 IU/ml (35 copies/ml) 30 1.1E8 IU/ml (170 6.4E8 copies/ml) IU/ml (fixed conversion factor to convert results to copies/ml) A-G; pre-core mutants 3 (negative, low & high positives) Yes, QS incorporated into each sample prior extraction ~ 5 hours

Assay time

A-E; limited F&G recovery 6 standard calibrators (standard curve) Yes, QS incorporated into each sample prior extraction ~ 8 hours

Technical Comparisons
sensitivity, samples detected below A HBM & CA HBM LOD

in upper end of linear range, in number of samples requiring dilution or repeat testing

10 9 8 7 6 5 4 3 2 1 0 0 1 2 3 4 5 6 7 8 9 10

COBAS AMPLICOR HBV MONITOR (1000-4E7 copies/ml) COBAS AMPLICOR HBV MONITOR (300-200,000 copies/ml) High Pure System/COBAS TaqMan HBV (170-6.4E8 copies/ml, 95% at 35 copies/mL)

High Pure System Viral Nucleic Acid Kit/COBAS TaqMan TM HCV Test Performance

HighPure System/COBAS TaqMan HCV Test Limit of Detection

Plasma (EDTA)
Input Titer IU/ml n Total Results hits %

Serum
Input Titer IU/ml n Total Results hits %

69 69 20.8 72 72 15.6 71 68 10.4 72 53 5.2 72 34 2.6 95% Hit Rate Concentration by Probit
95% confidence interval

100.0% 100.0% 95.8% 73.6% 47.2% 9.56 7.92 - 12.73

20.8 67 67 15.6 72 69 10.4 72 68 5.2 72 64 2.6 69 41 95% Hit Rate Concentration by Probit 95% confidence interval

100.0% 95.8% 94.4% 88.9% 59.4% 10.27 8.08-14.86

High Pure System/COBAS TaqMan HCV Test Linearity/Dynamic Range Study - Serum
Serum Linearity Results: HCV Armored RNA
10.00 9.00 8.00

Log Observed

7.00 6.00 5.00 4.00 3.00 2.00 1.00 0.00

y = 0.9377x + 0.2424 R2 = 0.9984 Levels 30-1e9 Levels 10 Linear (Levels 30-1e9)

0.00

2.00

4.00

6.00

8.00

10.00

Log Nominal

HPS/CTM HCV linear range = 30 - 2E8 IU/ml

High Pure System/COBAS TaqMan HCV Test Linearity/Dynamic Range Study Serum
Serum Linearity Results Clinical Panel
7.0 6.0

y = 1.0043x + 0.1082 R 2 = 0.9967

Log Titer - Observed

5.0 4.0 3.0 2.0 1.0 0.0 0.0 1.0 2.0 3.0 4.0 Log Titer - Nominal 5.0 6.0 7.0 Levels 1.48e1 - 1.32e6 Linear (Levels 1.48e1 - 1.32e6)

High Pure System/COBAS TaqMan HCV Test Specificity

Specimen Type Number of Specimens HPS/CTM HCV Test Results Tested Not Detected Reactive % Specificity
Unique Negative EDTA Plasma 301 301 0 100

Serum

303

303

100

Technical Comparisons
AMPLICOR HCV MONITOR (600-850,000 IU/mL/mL) COBAS AMPLICOR HCV MONITOR (600-500,000 IU/mL/mL) High Pure System/COBAS TaqMan Test HCV (30->2E8 IU/mL, LOD = 95% at 10 IU/mL)

sensitivity, samples detected below CA HCM LOD

in upper end of linear range, in number of samples requiring dilution or repeat testing

10 9 8 7 6 5 4 3 2 1 0 0 1 2 3 4 5 6 7 8 9 10

Comparison
CA HCM v. HPS/CTM HCV
Test/Test Parameters
Tests/kit (Minimum batch size) Limit of Quantitation (LOQ) Limit of Detection (LOD) Linear Range Specificity Reportable Units Genotype Inclusivity Sample Types Sample Volume # of controls Quantitation Standard Assay Time

CA HCM
48 (12) 600 IU/mL 600 IU/mL 600-700,000 copies/ml >99.99% IU/mL 1-6 Serum, plasma 200 uL 3 (High, Low, Neg) Yes ~8.5 hours

HPS/CTM HCV
48 (12) 30 IU/mL 10 IU/mL 30 >2E8 IU/mL >99.99% IU/mL 1-6 Serum, plasma 500 uL 3 (High, Low, Neg) Yes ~ 5.0 hours

High Pure System/COBAS TaqMan HCV Test Correlation to Bayer Versant HCV bDNA 3.0
Platform Correlation Study
1 22 Specimens (99 Serum, 23 EDTA Plasma) with VERSANT Titers >61 5 IU/mL 1 6 Specimens (Serum) with VERSANT Titers <61 5 IU/mL
10

8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 y = 1.0525x + 0.0195 R2 = 0.8975 Linear (615 IU/mL < Titer < 7.7E+6 IU/mL) 615 IU/mL > Titer > 1 IU/mL 615 IU/mL < Titer < 7.7E+6 IU/mL

COBAS TaqMan HCV Test Log Titer, IU/mL

VERSANT HCV RNA 3.0 Assay Log10 Titer, IU/mL

Comparison
HPS/CTM HCV vs. Bayer VERSANT bDNA
Test/Test Parameters
Tests/kit (Minimum batch size) Limit of Quantitation (LOQ) Limit of Detection (LOD) Linear Range Specificity Reportable Units Genotype Inclusivity Sample Types Sample Volume # of controls Quantitation Standard Assay Time

HPS/CTM HCV
48 (12) 30 IU/mL 10 IU/mL 30 >2E8 IU/mL >99.99% IU/mL 1-6 Serum, plasma 500 uL 3 (High, Low, Neg) Yes ~ 5.0 hours

VERSANT bDNA
96 (12) 615 IU/mL 1000 IU/mL 615-7.69E7 IU/mL 98.8% IU/mL 1-6 Serum, plasma 50 uL 12 (9 standards, 3 controls) No ~20-24 hours

High Pure System Viral Nucleic Acid Kit/COBAS TaqMan TM HIV-1 Test Performance

HighPure/COBAS TaqMan 48 HIV-1 Test: Linear Range

HighPure-COBAS TaqMan HIV-1 test, Result Log10 (HIV-1 RNA copies/mL)
8 7 6 5 4 3 2 1 0 1 2 3 4 5 6 7 8

Data support: 37 cp/mL to 1.03E7 cp/mL Claim: 40 cp/mL - 1E7 cp/mL

y = 1.0235x + 0.00008 R2 = 0.986

Nominal Concentration Log10 (HIV-1 RNA copies/mL)

HighPure/COBAS TaqMan 48 HIV-1 Test: Specificity

Specimen Type Kit Lot Number of Specimens Tested

Unique Negative EDTA Plasma SLD-1 100

COBAS TaqMan HIV-1 Results Not Detected Reactive % Specificity

100 0 100

HighPure/COBAS TaqMan 48 HIV-1 Test: Interference of HIV Antiviral Drugs

Effect of antiviral drugs on the High Pure System Viral nucleic Acid/COBAS TaqMan HIV-1 Test (Drugs: average of 3 replicates; TC: average of 28 replicates)
100000.0
Log10 HIV-1 RNA copies/mL

10000.0 1000.0

1X Cmax
100.0 10.0 1.0
Zi ag en Vi re ad Cr ix i va Fo n rto va se No rv ir Vi ra ce Ag pt en er as e Ka le tra Vi ra m un e Su st iv a Cy to ve ne Va l cy te Zo vi r ax Va l tr ex In tro n A Ro fe Fu ro n ze A on Te ( Tst 20 co ) nt ro l( TC ) Re tro vi r Ep v ir Hi v id Ze rit

3X Cmax

Antiviral Drugs

Roche HIV-1 MONITOR Tests Comparison

CAP/CA HIV-1 MONITOR v1.5 Extraction Procedure Amplification Dynamic Range Lower Quantitation Limit (LOQ) Detection Rates at LOQ Specificity Units Reported HIV-1 Subtypes Reproducibility Contamination Control Specimen Type Sample Volume Numbers of controls Number of Standards Hands-on-Time Time-to-Result Internal Control Specimen Preparation Automation Probe Capture AMPLICOR RT-PCR US: 50 - 100,000 Std: 500 - 1,000,000 50 copies/mL 95% 99.85% Copies/mL A-H 25 - 35% Amperase EDTA Plasma US: 700 L Std: 200 L 3 1 IQS 30 minutes 7 hours Yes COBAS AmpliPrep COBAS AMPLICOR HPS/CTM HIV-1 Silicon columns using HighPure Real time PCR 40 - 10,000,000 40 copies/mL 95% 100% Copies/mL A-H Amperase EDTA Plasma 500 L 3 1 IQS 1.5 hours 5 hours Yes HigPure COBAS TaqMan 48 CAP/CTM HIV-1 Automated Magnetic Beads Real Time PCR 40 - 10,000,000 40 copies/mL 95% 100% Copies/mL A-H Amperase EDTA Plasma 850 L 3 1 IQS 30 minutes 6 hours Yes COBAS AmpliPrep COBAS TaqMan 48/96

TaqMan Assay Principle: 5Nuclease

Additional Instrumentation required

Water bath - 50 C Sigma Bench Top Centrifuge - 4600 g
Centrifuge art 10732 Swing Out Rotor art.11118

Check Price at local Sigma Distributors (Export)

HPS/CTM HCV Test Performance Characteristics Sensitivity: Dynamic Range: 10 IU/ml 30 - 2E8 IU/ml

Specificity:
Genotype inclusivity:

>99.99%
1-6

Sample Input: or Serum

Time to Result:

500 l EDTA Plasma

~ 5 hours

TaqMan Reagents: Performance for HBV (report in IU/ml)

Dynamic: 50- 10E9 c/ mL OR Range 10- 200M IU/ mL Sensitivity: 50 copies or 10 IU Subtype: A- G Total: 200 Precision 2 x 10E3 2 x 10E4 2 x 10E5 2 x 10E6 2 x 10E7 60% 30% 30% 30% 30% 40%

TaqMan Reagents: Performance for HIV-1

Dynamic Range: 30- 10E7 c/ mL Sensitivity: 30 copies Subtype: all group- M Precision >10E3 25%

Future Parameters COBAS TaqMan 48 Analyzer

CT CT/NG CMV HSV 1,2 FV Leiden Prothrombin (G20210A) Cytochrome P450 2C9
r

Advantages of the Taqman Tests

Single-tube amplification and real time detection: QS and Target Eliminates post-PCR manipulations Closed system: minimizes contamination Fewer reagents and disposables required Sensitive and probe-specific Broad dynamic range Walk-away system Small volume of tests/ day r