HBV Test v2.0 - 72

COBAS AmpliPrep/COBAS TaqMan
HBV Test, version 2.0

FOR IN VITRO DIAGNOSTIC USE.
HBV Test, v2.0
HBV V2.0
72 Tests
P/N: 04894570 190

Wash Reagent
PG WR
5.1 Liters
P/N: 03587797 190
INTENDED USE
The COBAS AmpliPrep/COBAS TaqMan HBV Test, version 2.0 (v2.0) is an in vitro nucleic acid
amplification test for the quantitation of Hepatitis B Virus (HBV) DNA in human plasma and serum, using
the COBAS AmpliPrep Instrument for automated specimen processing and the COBAS TaqMan
Analyzer or COBAS TaqMan 48 Analyzer for automated amplification and detection.
This test is intended for use as an aid in the management of patients with chronic HBV infection
undergoing anti-viral therapy. The assay can be used to measure HBV DNA levels at baseline and during
treatment to aid in assessing response to treatment. The results from the COBAS AmpliPrep/COBAS
TaqMan HBV Test, v2.0 must interpreted within the context of all relevant clinical and laboratory findings.
The COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 is not intended for use as a screening
test for the presence of HBV in blood or blood products or as a diagnostic test to confirm the
presence of HBV infection.
SUMMARY AND EXPLANATION OF THE TEST
Hepatitis B Virus (HBV) is one of several viruses known to cause viral hepatitis. Over two billion people
throughout the world have been infected with HBV and over 350 million of them are chronically infected
carriers1,2. Chronic carriers are at high risk of long term complications of infection, including chronic
hepatitis, cirrhosis and hepatocellular carcinoma3,4,5. Serologic markers are commonly used as diagnostic
and/or prognostic indicators of acute or chronic HBV infection. The most common marker of HBV infection
is the presence of HBV surface antigen (HBsAg). Although carriers may clear HBsAg and develop antibody
to HBsAg, there appears to still be a risk of serious liver complications later in life6,7. HBV e antigen
(HBeAg) is generally used as a secondary marker to indicate active HBV replication associated with
progressive liver disease. Failure to clear HBeAg appears to increase the risk of end stage liver disease8,9.
Variant strains of HBV can either produce HBeAg that is not detectable in serum or the strain can lose the
ability to make HBeAg even when an active infection is present10. Therefore, using this marker to monitor
disease progression may be of limited utility11. The ability to detect HBV DNA in serum has been reported
to have prognostic value for the outcome of acute and chronic HBV infections12-15. The methodology can
allow the detection of HBV DNA after HBsAg clearance16 or detection of HBV lacking serologic markers17.
However, a relationship between serologic markers and HBV DNA levels has not yet been established. The
efficacy of antiviral therapy used to treat patients with HBV can also be assessed by serologic markers or
by measurement of liver enzyme function. However, the most direct and reliable measurement of viral
replication is thought to be quantitation of HBV viral DNA in serum or plasma14,18-20. A rapid and sustained
drop in HBV DNA levels in patients receiving treatment with alpha-interferon, Lamivudine, Ganciclovir or
Adefovir Dipivoxil has been shown to be a predictive factor for a favorable treatment outcome11,21-25.
Monitoring of HBV DNA levels can predict the development of resistance to Lamivudine26. Therefore, a
quantitative test for the measurement of HBV DNA is a valuable tool that can be used in conjunction with
other serological markers in the management of HBV infection.
The Document Revision Information section is located at the end of this document.
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HBV DNA in plasma and serum can be quantitated by nucleic acid amplification technologies, such as the
Polymerase Chain Reaction (PCR)27-29. The COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 uses
PCR technology to achieve maximum sensitivity and dynamic range for the quantitative detection of HBV
DNA in EDTA anticoagulated plasma and serum.
PRINCIPLES OF THE PROCEDURE
The COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 is a nucleic acid amplification test for the
quantitation of Hepatitis B Virus (HBV) DNA in human plasma and serum. The COBAS
AmpliPrep/COBAS TaqMan HBV Test, v2.0 is based on two major processes: (1) specimen preparation
to isolate HBV DNA and (2) simultaneous PCR amplification27 of target DNA and detection of cleaved
dual-labeled oligonucleotide detection probe specific to the target.
The COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 permits automated specimen preparation followed
by automated PCR amplification and detection of HBV target DNA and HBV Quantitation Standard (QS) DNA.
The Master Mix reagent contains primer pairs and probes specific for both HBV DNA and HBV QS DNA. The
Master Mix has been developed to ensure equivalent quantitation of genotypes A through H of HBV. The
detection of amplified DNA is performed using a target-specific and a QS-specific dual-labeled oligonucleotide
probe that permit independent identification of HBV amplicon and HBV QS amplicon.
The quantitation of HBV viral DNA is performed using the HBV QS. It compensates for effects of inhibition
and controls the preparation and amplification processes, allowing a more accurate quantitation of HBV
DNA in each specimen. It is a non-infectious DNA construct that contains HBV sequences with identical
primer binding sites as the HBV target DNA and a unique probe binding region that allows HBV QS
amplicon to be distinguished from HBV amplicon.
The HBV QS is added to each specimen at a known copy number and is carried through the subsequent
steps of specimen preparation, simultaneous PCR amplification and detection of cleaved dual-labeled
oligonucleotide detection probes. The COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer
calculates the HBV DNA concentration in the test specimens by comparing the HBV signal to the HBV QS
signal for each specimen and control.
Target Selection
Selection of the target DNA sequence for HBV depends on identification of regions within the HBV genome
that show maximum sequence conservation among the various HBV genotypes. Generic silica-based specimen
preparation is used to capture the HBV DNA and HBV QS DNA and defined oligonucleotides are used as
primers in amplification of the HBV DNA and HBV QS DNA. A target-specific and a QS-specific dual-labeled
oligonucleotide probe permit independent identification of HBV amplicon and HBV QS amplicon. Accordingly,
the appropriate selection of the primers and the dual-labeled oligonucleotide probe is critical to the ability of
the test to amplify and detect the HBV genotypes. The COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0
uses three amplification primers for PCR. A signal generating dual-labeled probe hybridizes to the anti-sense
strand and is cleaved by Z05 DNA polymerase during extension of the primers. The amplicon of genotypes A-F
and H consists of a sequence of 145 nucleotides. Genotype G amplicon, however, consists of a sequence of
181 nucleotides. A 36 base insertion within the single-stranded highly conserved pre-Core/Core region of the
HBV genome results in a larger amplicon for genotype G31.
Specimen Preparation
The COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 utilizes automated specimen preparation on the
COBAS AmpliPrep Instrument by a generic silica-based capture technique. The procedure processes 500 L
of plasma or serum. The HBV virus particles are lysed by incubation at elevated temperature with a protease
and chaotropic lysis/binding buffer that releases nucleic acids and protects the released HBV DNA from
DNases in plasma and serum. Protease and a known number of HBV QS DNA molecules are introduced into
each specimen along with the lysis reagent and magnetic glass particles. Subsequently, the mixture is
incubated and the HBV DNA and HBV QS DNA are bound to the surface of the glass particles. Unbound
substances, such as salts, proteins and other cellular impurities, are removed by washing the magnetic glass
particles. After separating the beads and completing the washing steps, the adsorbed nucleic acids are eluted
at elevated temperature with an aqueous solution. The processed specimen, containing the magnetic glass
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particles as well as released HBV DNA and HBV QS DNA, is added to the amplification mixture and transferred
to the COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer.
PCR Amplification
The PCR amplification reaction is performed with the thermostable recombinant enzyme Thermus specie
2+
Z05 DNA Polymerase (Z05). In the presence of manganese (Mn ) and under the appropriate buffer
conditions, Z05 has DNA polymerase activity32, 33. This allows PCR amplification to occur together with
real-time detection of the amplicon.
Processed specimens are added to the amplification mixture in amplification tubes (K-tubes) in which PCR
amplification occurs. In the presence of Mn2+ and excess deoxynucleotide triphosphates (dNTPs),
including deoxyadenosine, deoxyguanosine, deoxycytidine and deoxyuridine triphosphates, Z05 DNA
polymerase extends the annealed primers forming a DNA strand.
Target Amplification
The Thermal Cycler in the COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer heats the
reaction mixture to denature the double-stranded DNA and expose the specific primer target sequences
on the HBV circular DNA genome and the HBV QS DNA. As the mixture cools, the primers anneal to the
target DNA. The thermostable Thermus specie Z05 DNA Polymerase (Z05) in the presence of Mn2+ and
excess deoxynucleotide triphosphates (dNTPs), including deoxyadenosine, deoxyguanosine, deoxycytidine
and deoxyuridine (in place of thymidine), extends the annealed primers along the target template to
produce a double-stranded DNA molecule termed an amplicon. The COBAS TaqMan Analyzer or
COBAS TaqMan 48 Analyzer automatically repeats this process for a designated number of cycles, with
each cycle intended to double the amount of amplicon DNA. The required number of cycles is
preprogrammed into the COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer. Amplification
occurs only in the region of the HBV genome between the primers; the entire HBV genome is not amplified.
Selective Amplification
Selective amplification of target nucleic acid from the specimen is achieved in the COBAS
AmpliPrep/COBAS TaqMan HBV Test, v2.0 by the use of AmpErase (uracil-N-glycosylase) enzyme and
deoxyuridine triphosphate (dUTP). The AmpErase enzyme recognizes and catalyzes the destruction of DNA
strands containing deoxyuridine34, but not DNA containing deoxythymidine. Deoxyuridine is not present in
naturally occurring DNA, but is always present in amplicon due to the use of deoxyuridine triphosphate as one
of the dNTPs in the Master Mix reagent; therefore, only amplicon contains deoxyuridine. Deoxyuridine renders
contaminating amplicon susceptible to destruction by the AmpErase enzyme prior to amplification of the target
DNA. Also, any nonspecific product formed after initial activation of the Master Mix by manganese is destroyed
by the AmpErase enzyme. The AmpErase enzyme, which is included in the Master Mix reagent, catalyzes the
cleavage of deoxyuridine-containing DNA at the deoxyuridine residues by opening the deoxyribose chain at the
C1-position. When heated in the first thermal cycling step, the amplicon DNA chain breaks at the position of
the deoxyuridine, thereby rendering the DNA non-amplifiable. The AmpErase enzyme remains inactive for a
prolonged period of time once exposed to temperatures above 55C, i.e. throughout the thermal cycling steps,
and therefore does not destroy target amplicon formed during amplification.
Detection of PCR Products in a COBAS TaqMan Test
The COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 utilizes real-time27,28 PCR technology. The use
of dual-labeled fluorescent probes allows for real-time detection of PCR product accumulation by
monitoring of the emission intensity of fluorescent reporter dyes released during the amplification process.
The probes consist of HBV and HBV QS-specific oligonucleotide probes with a reporter dye and a
quencher dye. In the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 the HBV and HBV QS probes
are labeled with different fluorescent reporter dyes. When these probes are intact, the fluorescence of the
reporter dye is suppressed by the proximity of the quencher dye due to Frster-type energy transfer
effects. During PCR, the probe hybridizes to a target sequence and is cleaved by the 5' 3' nuclease
activity of the thermostable Z05 DNA polymerase. Once the reporter and quencher dyes are released and
separated, quenching no longer occurs, and the fluorescent activity of the reporter dye is increased. The
amplification of HBV DNA and HBV QS DNA are measured independently at different wavelengths. This
process is repeated for a designated number of cycles, each cycle effectively increasing the emission
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intensity of the individual reporter dyes, permitting independent identification of HBV and HBV QS DNA.
The PCR cycle where a growth curve starts exponential growth is related to the amount of starting material
at the beginning of the PCR.
Fundamentals of COBAS TaqMan Test Quantitation
The COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 is quantitative over a very wide dynamic range
since the monitoring of amplicon is performed during the exponential phase of amplification. The higher
the HBV titer of a specimen, the earlier the fluorescence of the reporter dye of the HBV probe rises above
the baseline fluorescence level (see Figure 1). Since the amount of HBV QS DNA is constant between all
specimens, the fluorescence of the reporter dye of the HBV QS probe should appear at the same cycle for
all specimens (see Figure 2). In specimens where the QS fluorescence is affected, the concentration is
adjusted accordingly. The appearance of the specific fluorescent signals is reported as a critical threshold
value (Ct). The Ct is defined as the fractional cycle number where reporter dye fluorescence exceeds a
predetermined threshold (the Assigned Fluorescence Level), and starts the exponential growth phase of
this signal (see Figure 3). A higher Ct value indicates a lower titer of initial HBV target material. A 2-fold
increase in titer correlates with a decrease of 1 Ct for target HBV DNA, while a 10-fold increase in titer
correlates with a decrease of 3.3 Ct.
Figure 1 shows the target growth curves for a dilution series spanning a 5-log10 range. As the
concentration of the virus increases, the growth curves shift to earlier cycles. Therefore, the leftmost
growth curve corresponds to the highest viral titer level, whereas, the rightmost growth curve corresponds
to the lowest viral titer level.
Figure 1
Target Growth Curves for a Dilution Series of Virus Spanning a 5-Log10 Range
9.00
8.00
Normalized Fluorescence
7.00
6.00
5.00
4.00
Highest Titer
3.00
2.00
1.00
Lowest Titer
0.00
-1.00
0
10
20
30
40
50
Cycle Number
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Figure 2 shows the Quantitation Standard growth curves for specimens from a viral dilution series that
spans a 5-log10 range. The amount of Quantitation Standard added to each specimen is constant for each
reaction. The Ct value of the Quantitation Standard is similar regardless of the viral titer.
Figure 2
Quantitation Standard Growth Curves for a Dilution Series of Virus Spanning a 5-Log10 Range
35.00
30.00
Lowest Titer
25.00
20.00
15.00
10.00
5.00
Highest Titer
0.00
-5.00
0
10
20
30
40
50
Cycle Number
Figure 3 provides an example of how the fluorescence values at every cycle are normalized for each
growth curve. The fractional cycle number (Ct) is calculated where the fluorescence signal crosses the
Assigned Fluorescence Level.
Figure 3
Fluorescence Values at Every Cycle are Normalized for Each Growth Curve
1
0.9
0.8
Ct Value = 27.1
Assigned Fluorescence
Level
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
-0.1
-0.2
19
20
21
22
23
24
25
26
27
28
29
Cycle Number
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HBV DNA Quantitation

The COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 quantitates HBV viral DNA by utilizing a second
target sequence (HBV Quantitation Standard) that is added to each test specimen at a known
concentration. The HBV QS is a non-infectious DNA construct, containing fragments of HBV sequences
with primer binding regions identical to those of the HBV target sequence. The HBV QS contains HBV
primer binding regions and generates an amplification product of the same length and base composition
as the HBV target DNA. The detection probe binding region of the HBV QS has been modified to
differentiate HBV QS amplicon from HBV target amplicon.
During the annealing phase of the PCR in the COBAS TaqMan Analyzer or COBAS TaqMan 48
Analyzer, the specimens are illuminated and excited by filtered light and filtered emission fluorescence
data are collected for each specimen. The readings from each specimen are then corrected for
instrumental fluctuations. These fluorescence readings are sent by the instrument to the AMPLILINK
software and stored in a database. Pre-Checks are used to determine if the HBV DNA and HBV QS DNA
data represent sets that are valid, and flags are generated when the data lie outside the preset limits. After
all Pre-Checks are completed and passed, the fluorescence readings are processed to generate Ct values
for the HBV DNA and the HBV QS DNA. The lot-specific calibration constants provided with the COBAS
AmpliPrep/COBAS TaqMan HBV Test, v2.0 are used to calculate the titer value for the specimens and
controls based upon the HBV DNA and HBV QS DNA Ct values. The COBAS AmpliPrep/COBAS
TaqMan HBV Test, v2.0 is standardized against the WHO International Standard for Hepatitis B Virus DNA
for Nucleic Acid Technology (NAT) Assays Testing (NIBSC 97/746)29 and titer results are reported in
International Units (IU/mL).
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REAGENTS
COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0
(P/N: 04894570 190)
HBV2 CS1
(HBV Magnetic Glass Particles Reagent Cassette)
Magnetic glass particles
93% Isopropanol
Xi
93% (w/w) Isopropanol
HBV V2.0
72 Tests
1 x 72 Tests
1 x 7.0 mL
Irritant
F
93% (w/w) Isopropanol
Highly
Flammable
HBV2 CS2
(HBV Lysis Reagent Cassette)
Sodium citrate dihydrate
42.5% Guanidine thiocyanate
< 5% Polydocanol
0.9% Dithiothreitol
Xn
42.5% (w/w) Guanidine thiocyanate
1 x 72 Tests
1x 78 mL
Harmful
N
< 5% (w/w) Polydocanol
Dangerous For
The Environment
HBV2 CS3
HBV Multi-Reagent Cassette containing:
1 x 72 Tests
Pase
(Proteinase Solution)
Tris buffer
< 0.05% EDTA
Calcium chloride
Calcium acetate
< 7.8% Proteinase
Glycerol
Xn
< 7.8% (w/w) Proteinase
1 x 3.8 mL
Harmful
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EB, v2.0
(Elution Buffer, v2.0)
Tris-base buffer
0.2% Methylparaben
1 x 8.1 mL
HBV2 CS4
HBV Test-Specific Reagent Cassette containing:
1 x 72 Tests
HBV QS, v2.0

(HBV Quantitation Standard, v2.0)
Tris-HCl buffer
EDTA
< 0.005% Poly rA RNA (synthetic)
< 0.001% Non-infectious, linearized, double-stranded
plasmid DNA containing an insert with HBV primer
binding sequences and a unique probe binding region
0.05% Sodium azide
1 x 3.6 mL
HBV MMX, v2.0

(HBV Master Mix, v2.0)
Tricine buffer
Potassium acetate
Potassium hydroxide
0.09% Sodium azide
Glycerol
< 0.04% dATP, dCTP, dGTP, dUTP
< 0.003% Upstream and downstream HBV primers
< 0.003% Oligonucleotide aptamer
< 0.003% Fluorescent-labeled oligonucleotide probes
specific for HBV and the HBV Quantitation
Standard
< 0.05% Z05 DNA Polymerase (microbial)
< 0.1% AmpErase (uracil-N-glycosylase) enzyme
(microbial)
1 x 3.4 mL
HBV Mn2+
(HBV Manganese Solution)
< 0.5% Manganese acetate
Glacial acetic acid
0.09% Sodium azide
1 x 19.8 mL
HBV H(+)C, v2.0

(HBV High Positive Control, v2.0)
< 0.001% Linearized, double-stranded plasmid DNA
containing HBV sequences
Negative Human Plasma, non-reactive by tests for
antibody to HCV, antibody to HIV-1/2, HIV p24
antigen and HBsAg; HIV-1 RNA, HCV RNA and HBV
DNA not detectable by PCR methods
0.1% ProClin 300 preservative
Xi
(3:1) mixture of 5-Chloro-2-methyl-2H-isothiazol-3-one and
2-Methyl-2H-isothiazol-3-one
6 x 1.0 mL
Irritant
R36/38: Irritating to eyes and skin
R43: May cause sensitization by skin contact
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HBV L(+)C, v2.0

(HBV Low Positive Control, v2.0)
< 0.001% Linearized, double-stranded plasmid DNA
containing HBV sequences
antibody to HCV, antibody to HIV-1/2, HIV p24 antigen
and HBsAg; HIV-1 RNA, HCV RNA and HBV DNA not
detectable by PCR methods

Xi
6 x 1.0 mL
Irritant
CTM () C
[COBAS TaqMan Negative Control (Human Plasma)]
antibody to HCV, antibody to HIV-1/2, HIV p24 antigen
and HBsAg; HIV-1 RNA, HCV RNA and HBV DNA not
detectable by PCR methods
Xi
6 x 1.0 mL
Irritant
HBV H(+)C, v2.0 Clip
(HBV High Positive Control, v2.0 Barcode Clip)
1 x 6 Clips
HBV L(+)C, v2.0 Clip

(HBV Low Positive Control, v2.0 Barcode Clip)
1 x 6 Clips
HBV () C Clip
(HBV Negative Control Barcode Clip)
1 x 6 Clips
COBAS AmpliPrep/COBAS TaqMan Wash Reagent

(P/N: 03587797 190)
1 x 5.1 L
PG WR
PG WR
(COBAS AmpliPrep/COBAS TaqMan Wash Reagent)
Sodium citrate dihydrate
< 0.1% N-Methylisothiazolone-HCl
WARNINGS AND PRECAUTIONS

A.
FOR IN VITRO DIAGNOSTIC USE.
B.
This test is for use with human serum or plasma collected in the anticoagulant EDTA.
C.
Do not pipette by mouth.
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D.
Do not eat, drink or smoke in laboratory work areas. Wear protective disposable gloves, laboratory
coats and eye protection when handling specimens and kit reagents. Wash hands thoroughly after
handling specimens and test reagents.
E.
Avoid microbial and ribonuclease contamination of reagents when removing aliquots from
control vials.
F.
The use of sterile disposable pipettes and DNase-free pipette tips is recommended.
G.
Do not pool controls from different lots or from different vials of the same lot.
H.
Do not mix reagent cassettes or controls from different kits.
I.
Do not open COBAS AmpliPrep cassettes and exchange, mix, remove or add bottles.
J.
Dispose of unused reagents, waste and specimens in accordance with country, federal, state and
local regulations.
K.
Do not use a kit after its expiration date.
L.
Material Safety Data Sheets (MSDS) are available on request from your local Roche office.
M.
Specimens and controls should be handled as if infectious using safe laboratory procedures such
as those outlined in Biosafety in Microbiological and Biomedical Laboratories35 and in the CLSI
36
Document M29-A3 . Thoroughly clean and disinfect all work surfaces with a freshly prepared
solution of 0.5% sodium hypochlorite in deionized or distilled water.
Note: Commercial liquid household bleach typically contains sodium hypochlorite at a
concentration of 5.25%. A 1:10 dilution of household bleach will produce a 0.5%
sodium hypochlorite solution.
N.
CAUTION: CTM () C, HBV L(+)C, v2.0 and HBV H(+)C, v2.0 contain Human Plasma derived
from human blood. The source material has been tested and found non-reactive for the presence of
Hepatitis B Surface Antigen (HBsAg), antibodies to HIV-1/2 and HCV, and HIV p24 Antigen. Testing
of Negative Human Plasma by PCR methods showed no detectable HIV-1 RNA, HCV RNA or HBV
DNA. No known test methods can offer complete assurance that products derived from human
blood will not transmit infectious agents. Therefore, all human sourced material should be
considered potentially infectious. CTM () C, HBV L(+)C, v2.0 and HBV H(+)C, v2.0 should be
handled as if infectious using safe laboratory procedures such as those outlined in Biosafety in
Microbiological and Biomedical Laboratories35 and in the CLSI Document M29-A336. Thoroughly
clean and disinfect all work surfaces with a freshly prepared solution of 0.5% sodium hypochlorite
in deionized or distilled water.
O.
HBV QS, v2.0, HBV Mn2+ and HBV MMX, v2.0 contain sodium azide. Sodium azide may react
with lead and copper plumbing to form highly explosive metal azides. While disposing of sodium
azide-containing solutions down laboratory sinks, flush the drains with a large volume of water to
prevent azide buildup.
P.
Wear eye protection, laboratory coats and disposable gloves when handling any reagent. Avoid
contact of these materials with the skin, eyes or mucous membranes. If contact does occur,
immediately wash with large amounts of water. Burns can occur if left untreated. If spills of these
reagents occur, dilute with water before wiping dry.
Q.
Do not allow HBV2 CS2 and liquid waste from the COBAS AmpliPrep Instrument, which contain
guanidine thiocyanate, to contact sodium hypochlorite (bleach) solution. These mixtures can
produce a highly toxic gas.
R.
When disposing of used COBAS AmpliPrep Sample Processing Units (SPUs), which contain
guanidine thiocyanate, avoid any contact with sodium hypochlorite (bleach) solution. These
mixtures can produce a highly toxic gas.
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STORAGE AND HANDLING REQUIREMENTS

A.
Do not freeze reagents or controls.
B.
Store HBV2 CS1, HBV2 CS2, HBV2 CS3 and HBV2 CS4 at 2-8C. Unused, these reagents are
stable until the expiration date indicated. Once used, these reagents are stable for 63 days at 2-8C
or until the expiration date, whichever comes first. HBV2 CS1, HBV2 CS2, HBV2 CS3 and HBV2
CS4 can be used for up to a maximum of 64 hours cumulative on board the COBAS AmpliPrep
Instrument. Reagents must be stored at 2-8C between instrument cycles.
C.
Store HBV H(+)C, v2.0; HBV L(+)C, v2.0 and CTM () C at 2-8C. The controls are stable until
the expiration date indicated. Once opened, any unused portion must be discarded.
D.
Store Barcode clips [HBV H(+)C, v2.0 Clip, HBV L(+)C, v2.0 Clip and HBV () C Clip] at 2-30C.
E.
Store PG WR at 2-30C. PG WR is stable until the expiration date indicated. Once opened, this
reagent is stable for 28 days at 2-30C or until the expiration date, whichever comes first.
MATERIALS PROVIDED
A.

(P/N: 04894570 190)
HBV V2.0
HBV2 CS1
(HBV Magnetic Glass Particles Reagent Cassette)
HBV2 CS2
(HBV Lysis Reagent Cassette)
HBV2 CS3
(HBV Multi-Reagent Cassette)
HBV2 CS4
(HBV Test-Specific Reagent Cassette)
HBV H(+)C, v2.0
(HBV High Positive Control, v2.0)
HBV L(+)C, v2.0
(HBV Low Positive Control, v2.0)
CTM () C
[COBAS TaqMan Negative Control (Human Plasma)]
HBV H(+)C, v2.0 Clip
(HBV High Positive Control, v2.0 Barcode Clip)
HBV L(+)C, v2.0 Clip
(HBV Low Positive Control, v2.0 Barcode Clip)
HBV () C Clip
(HBV Negative Control Barcode Clip)
B.
COBAS AmpliPrep/COBAS TaqMan Wash Reagent

(P/N: 03587797 190)
PG WR
PG WR
(COBAS AmpliPrep/COBAS TaqMan Wash Reagent)
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MATERIALS REQUIRED BUT NOT PROVIDED

Instrumentation and Software
COBAS AmpliPrep Instrument

COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer
Optional: Docking Station
Optional: cobas p 630 instrument
AMPLILINK Software revision 3.2 series or 3.3 series
Data Station for the AMPLILINK software, with printer
AMPLILINK Software v3.2 or v3.3 Series Manuals:

COBAS AmpliPrep instrument Instrument Manual For use with the COBAS TaqMan
analyzer, COBAS TaqMan 48 analyzer, COBAS AMPLICOR analyzer, or cobas p 630
instrument, and the AMPLILINK software, version 3.2 and 3.3 series
COBAS TaqMan analyzer (plus optional docking station) Instrument Manual For use with
the AMPLILINK software, version 3.2 and 3.3 series Application Manual
COBAS TaqMan 48 analyzer Instrument Manual For use with the AMPLILINK software,
version 3.2 and 3.3 series Application Manual
AMPLILINK Software Version 3.2 Series Application Manual For use with the COBAS
AmpliPrep Instrument, COBAS TaqMan Analyzer, COBAS TaqMan 48 Analyzer, and
COBAS AMPLICOR Analyzer
or
AMPLILINK software Version 3.3 Series Application Manual For use with COBAS
AmpliPrep instrument, COBAS TaqMan analyzer, COBAS TaqMan 48 analyzer,
COBAS AMPLICOR analyzer, and cobas p 630 instrument
Optional: cobas p 630 instrument Operators Manual Software Version 2.2
Disposables
Sample processing units: SPUs

Sample input tubes (S-tubes) with barcode clips
Racks of K-tips
K-tube Box of 12 x 96
OTHER MATERIALS REQUIRED BUT NOT PROVIDED
Sample Rack (SK 24 rack)

Reagent Rack
SPU rack
K-carrier
K-carrier Transporter
K-carrier rack
Pipettors with aerosol barrier or positive displacement DNase-free tips (capacity 1000 L)*
Disposable gloves, powderless
Vortex mixer
* Pipettors should be accurate within 3% of stated volume. Aerosol barrier or positive displacement
DNase-free tips must be used where specified to prevent specimen and amplicon cross-contamination.
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SPECIMEN COLLECTION, TRANSPORT AND STORAGE

Note: Handle all specimens and controls as if they are capable of transmitting infectious agents.
Note: This test has been validated for use with only human serum or plasma collected in EDTA
anticoagulant. Testing of other specimen types may result in inaccurate results.
A.
Specimen Collection
The COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 is for use with plasma or serum specimens.
Blood should be collected in sterile tubes using EDTA (lavender top) as the anticoagulant or in SST
Serum Separation Tubes.
Store whole blood at 2-25C for no longer than 24 hours. Separate plasma or serum from whole blood
within 24 hours of collection by centrifugation at 800-1600 x g for 20 minutes at room temperature
(15-30C). Transfer plasma or serum to a sterile polypropylene tube. Figures 4 and 5 show specimen
stability data from specimen stability studies performed with the COBAS AmpliPrep/COBAS TaqMan
HBV Test, v2.0.
Figure 4
HBV Stability in Whole Blood (Collected in EDTA Plasma Tubes)
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Figure 5
HBV Stability in Whole Blood (Collected in Serum Separator Tubes)
B.
Specimen Transport
Transportation of whole blood, plasma or serum must comply with country, federal, state and local
regulations for the transport of etiologic agents37. Whole blood must be transported at 2-25C and
centrifuged within 24 hours of collection. Plasma or serum may be transported at 2-8C or frozen at -20C
to -80C.
C.
Specimen Storage
Plasma or serum specimens may be stored at 25-30C for up to 3 days, at 2-8C for up to 7 days or frozen
at -20C to -80C for at least six weeks. It is recommended that specimens be stored in 800-900 L
aliquots in sterile, 2.0 mL polypropylene screw-cap tubes (such as Sarstedt 72.694.006). Figures 6 and 7
show the specimen stability data from specimen storage studies performed with the COBAS
AmpliPrep/COBAS TaqMan HBV Test, v2.0.
Figure 6
HBV Stability in EDTA-Plasma
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Figure 7
HBV Stability in Serum
Plasma and serum specimens may be frozen and thawed up to five times without a loss of HBV DNA.
Figures 8 and 9 show the data from a freeze-thaw study performed with the COBAS AmpliPrep/COBAS
TaqMan HBV Test, v2.0
Figure 8
HBV Results after up to Five Freeze-Thaw Cycles (EDTA Plasma)
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Figure 9
HBV Results after up to Five Freeze-Thaw Cycles (Serum)
INSTRUCTIONS FOR USE

Note: For detailed operating instructions, a detailed description of the possible configurations,
printing results and interpreting flags, comments and error messages, refer to (1) the
COBAS AmpliPrep instrument Instrument Manual For use with the COBAS TaqMan
analyzer, COBAS TaqMan 48 analyzer, COBAS AMPLICOR analyzer, or cobas p 630
instrument, and the AMPLILINK software, version 3.2 and 3.3 series; (2) the COBAS
TaqMan analyzer (plus optional docking station) Instrument Manual For use with the
AMPLILINK software, version 3.2 and 3.3 series Application Manual; (3) the COBAS
TaqMan 48 analyzer Instrument Manual For use with the AMPLILINK software, version 3.2
and 3.3 series Application Manual; (4) the AMPLILINK Software Version 3.2 Series
Application Manual For use with the COBAS AmpliPrep Instrument, COBAS TaqMan
Analyzer, COBAS TaqMan 48 Analyzer, and COBAS AMPLICOR Analyzer or the
AMPLILINK software Version 3.3 Series Application Manual For use with COBAS
AmpliPrep instrument, COBAS TaqMan analyzer, COBAS TaqMan 48 analyzer,
COBAS AMPLICOR analyzer, and cobas p 630 instrument; (5) Optional: cobas p 630
instrument Operators Manual Software Version 2.2.
Batch Size
Each kit contains reagents sufficient for 72 tests, which may be performed in batches of 12 to 24 tests. At
least one of each control (CTM () C, HBV L(+)C, v2.0 and HBV H(+)C, v2.0) must be included in
each batch (see "Quality Control" section).
Workflow
The COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer run must be started within 120 minutes
following completion of specimen and control preparation.
Note: Do not freeze or store processed specimens and controls.
Specimen and Control Preparation
Note: If using frozen specimens, place the specimens at room temperature (15-30C) until
completely thawed and vortex for 3-5 seconds before use. Controls should be removed
from 2-8C storage and equilibrated to room temperature before use.
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COBAS AmpliPrep Instrument Set-up

Part A.
Maintenance and Priming
A1.
The COBAS AmpliPrep Instrument is ready for operation in stand-by mode.
A2.
Turn the Data Station for the AMPLILINK software ON. Prepare the Data Station as follows:
a.
Log onto the Windows XP operating system.
b.
Double click the AMPLILINK software icon.
c.
Log onto AMPLILINK software by entering the assigned User ID and password.
A3.
Check the supply of PG WR using the Status Screen and replace if necessary.
A4.
Perform all Maintenance that is listed in the Due Tab. The COBAS AmpliPrep Instrument will
automatically prime the system.
Part B.
Loading of Reagent Cassettes
Note: All reagent cassettes should be removed from 2-8C storage, immediately loaded onto the
COBAS AmpliPrep Instrument and allowed to equilibrate to ambient temperature on the
instrument for at least 30 minutes before the first specimen is to be processed. Do not let reagent
cassettes come to ambient temperature outside the instrument as condensation may form on the
barcode labels. Do not wipe off condensation if it appears on the barcode labels.
B1.
Place HBV2 CS1 onto a reagent rack. Place HBV2 CS2, HBV2 CS3 and HBV2 CS4 onto a
separate reagent rack.
B2.
Load the reagent rack containing HBV2 CS1 onto rack position A of the COBAS AmpliPrep
Instrument.
B3.
Load the reagent rack containing HBV2 CS2, HBV2 CS3 and HBV2 CS4 onto rack position B, C,
D or E of the COBAS AmpliPrep Instrument. (See Table 1 for additional information).
Part C.
Loading of Disposables
Note: Determine the number of COBAS AmpliPrep reagent cassettes, Sample Processing Units
(SPUs), Input Sample tubes (S-tubes), K-tips and K-tubes needed. One SPU, one Input Stube, one K-tip and one K-tube are needed for each specimen or control.
Multiple workflows for use of the COBAS AmpliPrep Instrument with the COBAS TaqMan Analyzer or
COBAS TaqMan 48 Analyzer are possible. For reference, see Table 1 below. Depending on the workflow
used, load the appropriate number of reagent cassette racks, sample racks with Input S-tubes, SPU racks,
K-tip racks, K-tube racks and K-carriers on K-carrier racks onto the respective rack positions of the
COBAS AmpliPrep Instrument (see Table 1 for additional information).
C1.
Place the SPUs in the SPU rack(s) and load the rack(s) onto rack position J, K or L of the COBAS
AmpliPrep Instrument.
C2.
Depending on the workflow used, load full K-tube rack(s) onto rack position M, N, O or P of the
COBAS AmpliPrep Instrument.
C3.
Load full K-tip rack(s) onto rack position M, N, O or P of the COBAS AmpliPrep Instrument.
C4.
For workflow 3 using the COBAS TaqMan 48 Analyzer, load K-carriers on K-carrier rack(s) onto
rack position M & N, or O & P of the COBAS AmpliPrep Instrument.
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Table 1
Possible Workflows for using COBAS AmpliPrep Instrument with
COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer
Workflow
COBAS
AmpliPrep
Instrument
plus
Docking Station
plus
COBAS TaqMan
Analyzer
COBAS
AmpliPrep
Instrument
plus
COBAS TaqMan
Analyzer
COBAS
AmpliPrep
Instrument
plus
COBAS TaqMan
48 Analyzer(s)
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Transfer Mode to
COBAS TaqMan
Analyzer or COBAS
TaqMan 48 Analyzer
Automated transfer of
K-carrier
Manual transfer of
K-tubes via sample
rack(s) onto COBAS
TaqMan Analyzer
Manual transfer of
K-carrier via
K-carrier rack(s)
onto COBAS
TaqMan 48
Analyzer
18
Racks, Carriers and

Disposables
K-tubes in full K-tube racks
K-tips in full K-tip racks
Input S-tubes containing
specimens and controls on
sample racks
SPUs in SPU racks
CS1 on Cassette rack
CS2, CS3, CS4 on Cassette
rack
K-tubes in full K-tube racks
sample racks
SPUs in SPU racks
rack
After specimen processing is
finished:
K-tubes on sample racks
(ready for manual transfer)
K-tubes on sample racks
sample racks
SPUs in SPU racks
rack
Empty barcoded K-carrier on
K-carrier rack
After specimen processing is
finished:
K-tubes in K-carrier on Kcarrier rack
Position on
COBAS
AmpliPrep
Instrument
MP
MP
FH
JL
A
BE
MP
MP
FH
JL
A
BE
Same as
above
(F H)
F-H
M-P
F-H
J-L
A
B-E
M-P
Same as
above
(M P)
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Part D.
Ordering and Loading of Specimens
D1.
Prepare sample racks as follows: Attach a barcode label clip to each sample rack position where a
specimen (S-tube) is to be placed. Attach one of the specific barcode label clips for the controls
[CTM () C, HBV L(+)C, v2.0 and HBV H(+)C, v2.0] to each sample rack position where the
controls (S-tube) are to be placed. The CTM () C must be placed in the sample rack position
following the last clinical specimen or positive control [HBV L(+)C, v2.0 or HBV H(+)C, v2.0] in
each batch of 12 to 24 tests. The barcode label clips for controls should have the same control lot
number as the lot number on the control vials in the kit. Take care in assigning the right control to
the position with the appropriate control barcode clip. Place one Input S-tube into each position
containing a barcode label clip.
D2.
Using the AMPLILINK software, create specimen orders for each specimen and control in the
Orders window Sample folder. Select the appropriate test file and complete by saving.
D3.
Assign specimen and control orders to sample rack positions in the Orders window Sample Rack
folder. The sample rack number must be for the rack prepared in Step D1.
D4.
Print the Sample Rack Order report to use as a worksheet.
D5.
Prepare specimen and control racks in the designated area for specimen and control addition as
follows: Vortex each specimen and control [CTM () C, HBV L(+)C, v2.0 and HBV H(+)C, v2.0]
for 3 to 5 seconds. Avoid contaminating gloves when manipulating the specimens and controls.
D6.
Transfer 650 L of each specimen and control [CTM () C, HBV L(+)C, v2.0 and HBV H(+)C,
v2.0] to the appropriate barcode labeled Input S-tube using a micropipettor with an aerosol barrier
or positive displacement DNase-free tip. Avoid transferring particulates and/or fibrin clots
from the original specimen to the Input S-tube. Specimens and controls should be transferred
to tube positions as assigned and recorded on the worksheet in Step D4. The barcode label clips for
controls should have the same control lot number as the lot number on the control vials in the kit.
Assign the right control to the position with the appropriate control barcode clip. Avoid contaminating the upper part of the S-tubes with specimens or controls.
D7.
If using the cobas p 630 instrument for preparation of specimens, refer to the cobas p 630
instrument Operators Manual.
D8.
For workflows 1 and 2, load the sample rack(s) filled with Input S-tubes onto rack positions F, G or
H of the COBAS AmpliPrep Instrument.
D9.
For workflow 3 using the COBAS TaqMan 48 Analyzer, load sample rack(s) with Input S-tubes
and K-tubes (one for each Input S-tube, loaded in the right position adjacent to Input S-tubes) onto
rack position F, G or H of the COBAS AmpliPrep Instrument.
Part E.
E1.
Part F.
Start of COBAS AmpliPrep Instrument Run

Start the COBAS AmpliPrep Instrument using the AMPLILINK software.
End of COBAS AmpliPrep Instrument Run and Transfer to COBAS TaqMan
Analyzer or COBAS TaqMan 48 Analyzer (for workflows 2 and 3 only)
F1.
Check for flags or error messages.
F2.
Remove processed specimens and controls from the COBAS AmpliPrep Instrument on either
sample racks (for COBAS TaqMan Analyzer without Docking Station) or K-carrier racks (for
COBAS TaqMan 48 Analyzer), depending on the workflow (for further details see Part G).
F3.
Remove waste from the COBAS AmpliPrep Instrument.
Note: Processed specimens and controls should not be exposed to light after completion of
specimen and control preparation.
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Amplification and Detection

COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer Set-up
The COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer run must be started within 120 minutes
following completion of specimen and control preparation.
Note: Do not freeze or store processed specimens and controls.
Part G.
G1.
Workflow 1:
Automated transfer of K-carrier via docking station to COBAS

TaqMan Analyzer. Manual intervention is unnecessary.
Workflow 2:
Manual transfer of K-tubes in sample rack(s) to COBAS TaqMan

Analyzer.
Workflow 3:
Manual transfer of K-carrier on K-carrier rack(s) to the COBAS

TaqMan 48 Analyzer. Manual transfer of K-carriers into COBAS
TaqMan 48 Analyzer using the K-carrier Transporter.
Part H.
H1.
Loading Processed Specimens
Depending on the workflow, perform the appropriate steps to transfer the K-tubes to the COBAS
TaqMan Analyzer or COBAS TaqMan 48 Analyzer:
Start of COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer Run
Start the COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer by one of the options
below depending on the workflow used:
Workflow 1:
No intervention necessary.
Workflow 2:
Automatic start of the COBAS TaqMan Analyzer after insertion of

sample rack(s).
Workflow 3:
Fill K-carrier with empty K-tubes if there are fewer than 6 K-tubes on
the K-carrier. Filling is guided by the AMPLILINK software. Open
thermal cycler cover, load K-carrier into thermal cycler and close lid.
Start the COBAS TaqMan 48 Analyzer run.
End of COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer Run
Part I.
I1.
At the completion of the COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer run, print
Results Report. Check for flags or error messages in the Result report. Specimens with flags and
comments are interpreted as described in the Results section. After acceptance, store data in archive.
I2.
Remove used K-tubes from the COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer.
RESULTS
The COBAS TaqMan Analyzer or the COBAS TaqMan 48 Analyzer automatically determines the HBV
DNA concentration for the specimens and controls. The HBV DNA concentration is expressed in
International Units (IU) /mL. The conversion factor between HBV copies/mL and HBV IU/mL is
5.82 copies/IU, using the WHO International Standard for Hepatitis B Virus DNA for Nucleic Acid
Technology (NAT) Assays Testing (NIBSC 97/746)29.
If needed, results can be converted manually to copies/mL using the conversion calculation as
follows:
HBV DNA concentration in IU/mL x 5.82 copies/IU = HBV DNA in copies/mL
Example: 1.23E+04 IU/mL x 5.82 copies/IU = 7.16E+04 copies/mL
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Note: The analytical measurement range of analyte values that can be directly measured on a
specimen without any dilution using the COBAS AmpliPrep/COBAS TaqMan HBV Test,
v2.0 is 20 to 1.7E+08 IU/mL.
Note: The clinical reportable range of analyte values that can be directly measured on a
specimen with a maximum dilution of one to ten using the COBAS AmpliPrep/COBAS
TaqMan HBV Test, v2.0 is 20 to 1.7E+09 IU/mL.
AMPLILINK Software:
Determines the Cycle Threshold value (Ct) for the HBV DNA and the HBV QS DNA.
Determines the HBV DNA concentration based upon the Ct values for the HBV DNA and
HBV QS DNA and the lot-specific calibration coefficients provided on the cassette barcodes.
Determines that the calculated IU/mL for HBV L(+)C, v2.0 and HBV H(+)C, v2.0 fall
within the fixed ranges.
Batch Validation AMPLILINK Version 3.2 Series

Check AMPLILINK software results window or printout for flags and comments to ensure that the batch is
valid. For control orders, a check is made to determine if the IU/mL value for the control is within its
specified range. If the IU/mL value for the control lies outside of its range, a FLAG is generated to show the
control has failed.
The batch is valid if no flags appear for any of the controls [HBV L(+)C, v2.0; HBV H(+)C, v2.0 and
CTM () C].
The batch is not valid if any of the following flags appear for the HBV Controls:
Negative Control
Flag
_N_NC_INVALID
Result
Invalid
Interpretation
An invalid result or a valid result that was not
negative for HBV target
HBV Low Positive Control, v2.0

Flag
Result
Interpretation
_L_LPCINVALID
<2.00E+01 IU/mL
Control below range
_L_LPCINVALID
Target Not Detected
Control below range
_L_LPCINVALID
A numeric titer
X.XXE+XX IU/mL
Control out of range
_L_LPCINVALID
> 1.70E+08 IU/mL
_L_LPCINVALID
Invalid
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Control above range

An invalid result
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HBV High Positive Control, v2.0

Flag
Result
Interpretation
_H_HPCINVALID
<2.00E+01 IU/mL
Control below range
_H_HPCINVALID
Target Not Detected
Control below range
_H_HPCINVALID
A numeric titer
X.XXE+XX IU/mL
Control out of range
_H_HPCINVALID
> 1.70E+08 IU/mL
_H_HPCINVALID
Invalid
Control above range

An invalid result
If the batch is invalid, repeat the entire batch including specimen and control preparation, amplification
and detection.
Batch Validation AMPLILINK Version 3.3 Series
Check AMPLILINK software results window or printout for flags and comments to ensure that the batch is
valid. For control orders, a check is made to determine if the IU/mL value for the control is within its
specified range. If the IU/mL value for the control lies outside of its range, a FLAG is generated to show the
control has failed.
The batch is valid if no flags appear for any of the controls [HBV L(+)C, v2.0; HBV H(+)C, v2.0 and
CTM () C].
The batch is not valid if any of the following flags appear for the HBV Controls:
Negative Control:
Flag
Result
Interpretation
NC_INVALID
Invalid
An invalid result or a valid result that was not negative for HBV target
HBV Low Positive Control, v2.0:

Flag
Result
Interpretation
LPCINVALID
Invalid
An invalid result or a control out of range
HBV High Positive Control, v2.0:

Flag
Result
Interpretation
HPCINVALID
Invalid
An invalid result or a control out of range
If the batch is invalid, repeat the entire batch including specimen and control preparation, amplification
and detection.
Interpretation of Results
For a valid batch, check each individual specimen for flags or comments on the result printout. Interpret
the results as follows:
A valid batch may include both valid and invalid specimen results depending on whether
flags and/or comments are obtained for the individual specimens.
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Specimen results are interpreted as follows:

Titer Result
Interpretation
Target Not Detected
Report results as "HBV DNA not detected".
<2.00E+01 IU/mL
Calculated IU/mL are below the Limit of Detection of the assay. Report
results as "HBV DNA detected, less than 20 HBV DNA IU/mL".
' 2.00E+01 IU/mL

and - 1.70E+08 IU/mL
Calculated results greater than or equal to 20 IU/mL and less than or equal
to 1.70E+08 IU/mL are within the Linear Range of the assay.
> 1.70E+08 IU/mL
Calculated IU/mL are above the range of the assay. Report results as
"greater than 1.70E+08 HBV DNA IU/mL". If quantitative results are
desired, the original specimen should be diluted with HBV-negative
human serum or EDTA plasma, depending on the matrix of the original
specimen, and the test repeated. Multiply the reported result by the
dilution factor.
Note: Specimens above the range of the assay that produce an Invalid result with a flag
"QS_INVALID" should not be reported as >1.70E+08 IU/mL. The original specimen should be
diluted with HBV-negative human serum or EDTA-plasma, depending on the matrix of the
original specimen, and the test repeated. Multiply the reported result by the dilution factor.
Note: Titer Result "Failed". Interpretation: Specimen is not correctly processed during specimen
preparation on the COBAS AmpliPrep Instrument.
Note: Titer Result "Invalid". Interpretation: An Invalid Result
QUALITY CONTROL
One COBAS TaqMan Negative Control, one HBV Low Positive Control, v2.0 and one HBV High Positive
Control, v2.0 must be included in each test batch. The batch is valid if no flags appear for any of the
controls [HBV L(+)C, v2.0; HBV H(+)C, v2.0 and CTM () C].
There are no requirements regarding the position of the positive controls [HBV L(+)C, v2.0 and HBV
H(+)C, v2.0] on the sample rack. However, the CTM () C must be placed in the sample rack position
following the last clinical specimen or positive control [HBV L(+)C, v2.0 or HBV H(+)C, v2.0] in each
batch of 12 to 24 tests.
Check the batch printout for flags and comments to ensure that the batch is valid.
Negative Control
The CTM () C must yield a "Target Not Detected" result. If the CTM () C is flagged as invalid, then the
entire batch is invalid. Repeat the entire process (specimen and control preparation, amplification and
detection). If CTM () C is consistently invalid in multiple batches, contact your local Roche office for
technical assistance.
Positive Controls
The fixed titer ranges for HBV L(+)C, v2.0 and HBV H(+)C, v2.0 are provided on the COBAS
AmpliPrep/COBAS TaqMan HBV Test, v2.0 reagent cassette barcodes.
The HBV DNA IU/mL for HBV L(+)C, v2.0 and HBV H(+)C, v2.0 should fall within their fixed titer
ranges. If one or both of the positive controls are flagged as invalid, then the entire batch is invalid. Repeat
the entire process (specimen and control preparation, amplification and detection). If the HBV DNA titer of
one or both of the positive controls is consistently outside the ranges in multiple batches, contact your
local Roche office for technical assistance.
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PROCEDURAL PRECAUTIONS
As with any test procedure, good laboratory technique is essential to the proper performance of this assay.
PROCEDURAL LIMITATIONS
1.
This test has been validated for use with human serum or plasma collected in EDTA anticoagulant.
Testing of other specimen types may result in inaccurate results.
2.
Reliable results are dependent on adequate specimen collection, transport, storage and processing
procedures.
3.
The presence of AmpErase enzyme in the COBAS AmpliPrep/COBAS TaqMan HBV Master Mix,
v2.0 reduces the risk of amplicon contamination. However, contamination from HBV positive
controls and clinical specimens can be avoided only by good laboratory practices and careful
adherence to the procedures specified in this Package Insert.
4.
Use of this product should be limited to personnel trained in the techniques of PCR.
5.
This product can only be used with the COBAS AmpliPrep Instrument and the COBAS TaqMan
Analyzer or COBAS TaqMan 48 Analyzer.
6.
Though rare, mutations within the highly conserved region of the viral genome covered by the
COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 primers and/or probe may result in the
under-quantitation of or failure to detect the virus.
7.
Detection of HBV DNA is dependent on the number of virus particles present in the specimen and
may be affected by specimen collection methods and patient factors, (i.e., age, presence of
symptoms, and/or stage of the infection).
8.
Due to inherent differences between technologies, it is recommended that, prior to switching from
one technology to the next, users perform method correlation studies in their laboratory to quantify
technology differences.
INTERFERING SUBSTANCES
Elevated levels of triglycerides, bilirubin and albumin in specimens have been shown not to interfere with the
quantitation of HBV DNA by this test. Elevated levels of hemoglobin in specimens up to a concentration of
250 mg/dL have been shown not to interfere with the quantitation of HBV DNA by this test.
The following drug compounds tested at 3 times of the Peak Plasma Level (Cmax) have been shown not to
interfere with the quantitation of HBV DNA by the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0:
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Nucleotide DNA Polymerase Inhibitors
Nucleoside Reverse Transcriptase and

DNA Polymerase Inhibitors
Tenofovir
Adefovir dipivoxil
Lamivudine
Telbivudine
Zidovudine
Stavudine
Abacavir
Didanosine
Entecavir
HIV Protease Inhibitors
Non-nucleoside HIV Reverse Transcriptase

Inhibitors
Indinavir
Nevirapine
Saquinavir
Efavirenz
Ritonavir
Amprenavir
HIV Fusion Inhibitors
Lopinavir/Ritonavir
Enfurvitide
Immune Modulators
Antidepressants
Interferon alpha-2a
Paroxetine HCl
Ribavirin
Fluoxetine
Interferon alpha-2b
Sertraline
Peginterferon alfa-2a
Compounds for Treatment of Herpes

Viruses
Peginterferon alfa-2a + Ribavirin

Interferon alpha-2b + Ribavirin
Ganciclovir
Peginterferon alfa-2b
Valganciclovir HCl
Acyclovir
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NON-CLINICAL PERFORMANCE EVALUATION

A.
Limit of Detection Using the WHO International Standard
The limit of detection of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 was determined for
HBV DNA by using dilutions of the WHO International Standard for Hepatitis B Virus DNA for Nucleic Acid
Technology (NAT) Assays Testing (NIBSC 97/746)29, genotype A in HBV-negative human EDTA plasma or
serum. Three dilution series were analyzed for each matrix. A total of 72 replicates per concentration were
tested for each matrix type.
The concentration of HBV DNA that can be detected with a positivity rate of greater than 95% as determined
by PROBIT Analysis, is 9 IU/mL for EDTA plasma (see Table 2) and 19 IU/mL for serum (see Table 3).
Table 2
Limit of Detection in EDTA Plasma of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0
using the WHO International Standard
Nominal Input
(HBV DNA IU/mL)
30
20
10
5
2.5
PROBIT 95% Hit Rate
No. Replicates
No. Positives
Positivity Rate
72
72
100%
71
71
100%
72
68
94%
72
63
88%
72
38
53%
9 IU/mL (95% confidence range: 6.8 12.1 IU/mL)
Table 3
Limit of Detection in Serum of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0
using the WHO International Standard
Nominal Input
(HBV DNA IU/mL)
30
20
10
5
2.5
PROBIT 95% Hit Rate
B.
No. Replicates
No. Positives
Positivity Rate
72
72
100%
72
70
97%
72
58
81%
72
29
40%
72
24
33%
19 IU/mL (95% confidence range: 11.0 107.1 IU/mL)
Limit of Detection Using Clinical Specimens Across All Genotypes
In addition, dilutions of clinical specimens in HBV-negative human EDTA plasma or serum representing
genotypes A-F and H and two quantified purified plasmid DNAs, one for genotype G and one for the precore mutant were analyzed with two kit lots of COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0
reagents. Twenty-four replicates per level, genotype, matrix and kit lot were tested.
The concentration of HBV DNA that can be detected with a hit rate of greater than 95% as determined by
PROBIT analysis, is 16.4 IU/mL combined for both matrices (see Table 4).
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Table 4
Limit of Detection in EDTA Plasma and Serum of the
COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 using

Genotype A-H and Pre-core Mutant
Genotype
A
B
C
D
E
F
G
H
Pre-core
Mutant
Plasma
95% PROBIT
hit rate
concentration
Lot 1
10.6
95% PROBIT
hit rate
concentration
Lot 2
<5*
Serum
Plasma
Serum
Plasma
10.4
5.6
5.8
5.8
5.5
5.3
16.4
5.8
Serum
Plasma
Serum
Plasma
14.8
5.9
5.6
13.1
10.9
10.4
9.8
6.1
Serum
Plasma
6.3
4.9
15.7
5.8
Serum
Plasma
6.0
12.3
5.9
5.9
Serum
Plasma
5.8
9.8
11.6
5.5
Serum
Plasma
Serum
11.6
10.4
5.8
11.2
11.2
12.2
Matrix
95% PROBIT
hit rate
concentration
(max.)
95%
PROBIT
confidence
interval
10.6
4.6 - 37.7
16.4
12.0 - 28.6
14.8
10.4 - 28.7
10.4
7.0 - 24.1
15.7
10.7 - 33.4
6.0
**
12.3
8.6 - 24.3
11.6
6.9 - 30.3
12.2
9.1 - 22.2
* No PROBIT analysis and 95% confidence interval could be determined due to 100% hit rate at all
concentration levels for lot 2
** No 95% confidence interval could be calculated by PROBIT analysis due to 100% hit rate of all tested
dilution levels
Note:
Stock concentrations for all tested specimens were determined via calibrator bracketing (ISO
11095:1996) by using the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0.
In order to avoid a potential bias, the concentrations of clinical genotype samples A-F and H
were verified with one or more of the following tests: Abbott RealTime HBV assay, the Siemens
VERSANT HBV DNA 3.0 assay and the COBAS TaqMan HBV Test For Use With The High
Pure System. Observed titers demonstrated a maximum deviation in quantitation of 0.3 Log10.
The concentrations for the two plasmids encompassing target regions for genotype G and the
pre-core mutant were verified via the PicoGreen method and accuracy of the assigned titer
was demonstrated in a dedicated comparability study.
05382874001-10EN
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C.
Precision
The precision of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 was determined by analysis of
serial dilutions of clinical HBV specimens (genotype A) in HBV-negative human EDTA plasma and serum.
The titer assignment of the clinical specimens (stock concentrations) was performed by a method that
ensures traceability to the WHO International Standard for Hepatitis B Virus DNA for Nucleic Acid
Technology (NAT) Assays Testing (NIBSC 97/746)29. 15 runs per matrix were performed, each consisting of
6 dilution levels and 3 replicates at each level. Each specimen was taken through the entire COBAS
AmpliPrep/COBAS TaqMan HBV Test, v2.0 procedure, including specimen preparation, amplification and
detection. Therefore, the precision reported here represents all aspects of the test procedure. The study
was performed using three lots of COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 reagents. The
results are shown in Table 5 (EDTA plasma) and Table 6 (serum).
Table 5
Precision of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 (EDTA Plasma Samples)
Lot 1
Titer
(IU/mL)
Total %CV
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
1.00E+07
26
17
12
8
11
14
Lot 2
Total SD
in log
0.12
0.07
0.05
0.04
0.05
0.06
Total %CV
28
17
11
8
11
12
Lot 3
Total SD
in log
0.13
0.07
0.05
0.04
0.05
0.06
Total SD
in log
0.11
0.09
0.06
0.06
0.05
0.08
Total %CV
25
20
14
13
12
18
Table 6
Precision of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 (Serum Samples)
D.
Lot 1
Titer
(IU/mL)
Total %CV
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
1.00E+07
27
13
11
10
11
15
Lot 2
Total SD
in log
0.13
0.06
0.05
0.04
0.05
0.07
Total %CV
22
15
10
12
12
15
Lot 3
Total SD
In log
0.11
0.06
0.04
0.05
0.05
0.07
Total %CV
28
17
14
11
11
13
Total SD
In log
0.13
0.08
0.06
0.05
0.05
0.06
Linear Range
As shown in Figure 10 and Figure 11, the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 was found
to give a linear response from 20 (Log10 = 1.30) HBV DNA IU/mL to 1.7E+08 (Log10 = 8.23) HBV DNA
IU/mL in both EDTA plasma and serum. The evaluation was performed according to CLSI Guideline EP6-A
using two lots of COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 reagents and serial dilutions of a
high titer HBV DNA (+) EDTA plasma or serum specimen. Thirteen concentration levels for EDTA plasma
and twelve levels for serum were tested, with 10 replicates per level.
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Figure 10
Linearity for the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0
in EDTA Plasma Specimens
10
8
7

HBV Test, v2.0 Results
Log10 (HBV DNA IU/mL)
6
5
Mean
4
3
2
y = 1.0183x - 0.0182
R = 0.9987
0
0
10
Nominal Concentration
Figure 11
Linearity for the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0
in Serum Specimens

HBV Test, v2.0 Results
9
8
7
6
5
Mean
4
3
2
y = 1.0202x - 0.1142
R = 0.9987
0
0
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E.
Genotype Titer Quantitation
To date, eight genotype categories have been proposed for HBV based on nucleotide divergence within
the genome of greater than 8%38, 39, 40. These genotypes are designated with capital alphabetical letters
41
from A through H. The HBV genotypes have characteristic geographic distributions .
The performance of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 on HBV genotypes was
evaluated by analysis of 25 purified, linearized and quantitated plasmid DNAs containing representative
sequence inserts from HBV genotypes A through H and the most common pre-core mutant (see Table 7).
The assignment of nominal concentrations to the plasmid stock solutions was performed by the PicoGreen
method. Each plasmid DNA was diluted to nominal concentrations of approximately 3.00E+03, 3.00E+05
and 3.00E+07 PicoGreen copies/mL in both EDTA plasma and serum. The concentrations (IU/mL) were
then determined by the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 using two reagent lots and
the results of all tested plasmids were compared.
The evaluation of all 25 plasmids analyzed by the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0
demonstrates equivalent quantification of all plasmids and genotypes for EDTA plasma and serum.
Table 7
Inclusivity Testing Typed Plasmid DNAs Tested
Plasmid Designation
p8423-c1
p1115-c1
p3952-c1
p4199-c2
p1764-c1
p1767-c1
p3958-c1
p830-c1
p3982-c1
p1786-c1
p11549-1
p3872-c1
p1103-c1
p3953-c2
p18-c1
p30893-5
p4244-c1
p3217-c1
p3963-c2
p9203-c1
p479-c1
p1009-c1
p00042975-4
pEFHBV20
pITmut1896
Genotype
A
E
F
G
H
Pre-core mutant
Parent Specimen Origin

India
Burundi
Cameroon
Norway
China
China
East Asia
Society Island
Vietnam
China
Bangladesh
Iran
Tunisia
North Africa
Sweden
Sweden
Denmark
Senegal
Nigeria
Colombia
Venezuela
Spain
United States
Nicaragua
Italy
Also, for eight of these plasmids, representing all genotypes, inclusivity was shown over the linear range
(Figure 12 and 13). For each plasmid, three concentration levels for EDTA plasma and serum were tested,
with 10 replicates per level.
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Figure 12
Performance of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0
on HBV Genotypes A through H in EDTA Plasma
0.4
Deviation
0.3
Genotype A
0.2
Genotype B
0.1
Genotype C
Genotype D
Genotype E
0.0
Genotype F
Genotype G
Genotype H
+/- 0.3 log
-0.1
-0.2
-0.3
-0.4
2.5
3.5
4.5
5.5
6.5
7.5
Figure 13
Performance of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0

on HBV Genotypes A through H in Serum
0.4
Deviation
0.3
Genotype A
0.2
Genotype B
Genotype C
Genotype D
Genotype E
Genotype F
0.1
0.0
-0.1
Genotype G
Genotype H
+/- 0.3 log
-0.2
-0.3
-0.4
2.5
3.5
4.5
5.5
6.5
7.5
F.
Clinical Specificity
The specificity of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 was determined by analysis of
HBV-negative EDTA plasma and serum from blood donors. A total of 647 individual EDTA plasma and 649
serum specimens were tested. The results of all specimens were negative for HBV DNA. Based on these
results, the specificity of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 is 100% with a
confidence limit of 99.54%.
05382874001-10EN
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G.
Cross Reactivity
The analytical specificity of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 was evaluated by
adding cultured organisms (viruses, bacteria, yeast) or DNA (HTLV-2, 2.5E+07 cp/PCR) into HBV-negative
human EDTA plasma (see Table 8).
None of the non-HBV organisms tested was positive for HBV DNA.
Table 8
Analytical Specificity Specimens
Virus
Adenovirus type 5
Cytomegalovirus
Epstein-Barr virus
Human Herpes Virus type 6
Herpes simplex virus type 1
Herpes simplex virus type 2
Human T-Cell Lymphotropic virus type 1
Human T-Cell Lymphotropic virus type 2
Influenza A
Hepatitis A virus
Hepatitis C virus
HIV-1
H.
Bacteria
Staphylococcus aureus
Propionibacterium acnes
Yeast
Candida albicans
Correlation of EDTA Plasma and Serum
The correlation of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 was tested by analysis of HBV
positive matched clinical EDTA plasma and serum specimens. 279 matched sets of specimens from HBV
infected patients were analyzed. The COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 results showed
high correlation between EDTA plasma and serum specimens (Figure 14).
Figure 14
Correlation of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0
on EDTA Plasma and Serum Specimens from HBV Infected Patients (n=279)
05382874001-10EN
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I.
Performance of COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 compared to the

COBAS AmpliPrep/COBAS TaqMan HBV Test
The performance of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 was compared to the
COBAS AmpliPrep/COBAS TaqMan HBV Test30 by analysis of HBV positive (' 54 IU/mL) clinical
specimens. Left over EDTA plasma specimens from routine diagnosis were analyzed at two external sites.
The specimen titers spread over the dynamic range of the COBAS AmpliPrep/COBAS TaqMan HBV
Test, v2.0. Linear regression analysis was performed on 275 valid samples (143 tested at one site and 132
at the other) (Figure 15).
Figure 15
and the COBAS AmpliPrep/COBAS TaqMan HBV Test

on EDTA Plasma Specimens from HBV Infected Patients (n=275)

Log10 HBV DNA Titer
9.00
8.00
7.00
6.00
y = 0.9958x + 0.0443
R 2 = 0.9743
5.00
4.00
3.00
2.00
1.00
0.00
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
COBASAmpliPrep/COBAS TaqMan HBV Test

Log10 HBV DNA Titer
J.
Method correlation
The performance of the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0 was compared to the
COBAS TaqMan HBV Test For Use with the High Pure System (HPS HBV, Figures 16 and 17), the
COBAS AMPLICOR HBV MONITOR Test (COBAS AMPLICOR HBV, Figures 18 and 19) and the
VERSANT HBV DNA 3.0 Assay (Figures 20 and 21) by regression analysis of undiluted clinically
characterized specimens from HBV infected patients. Specimens were obtained from Teragenix (Fort
Lauderdale, FL, USA). Regression analysis was performed for serum and EDTA-plasma specimens that
yielded results within the linear range.
05382874001-10EN
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Figure 16
and the COBAS TaqMan HBV Test For Use with the High Pure System

Plasma Specimen (Log10 Titer)
10.0
9.0
8.0
7.0
6.0
5.0
4.0
3.0
2.0
Y = 0.974 * X + 0.107
2
R = 0.964
1.0
0.0
0.0
2.0
4.0
6.0
8.0
10.0
High Pure System HBV

Plasm a Specim en (log10 Titer)
05382874001-10EN
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Figure 17
and the COBAS TaqMan HBV Test For Use with the High Pure System
on Serum Specimens from HBV Infected Patients (n=142)
05382874001-10EN
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Figure 18
and the COBAS AMPLICOR HBV MONITOR Test

5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
Y = 0.899 * X + 0.546
R 2 = 0.872
0.5
0.0
0.0
1.0
2.0
3.0
4.0
5.0
COBAS AMPLICOR HBV

05382874001-10EN
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Figure 19
and the COBAS AMPLICOR HBV MONITOR Test


Serum Specimen (Log 10 Titer)
Y = 0.911 * X + 0.461
2
R = 0.821
0
0
COBASAMPLICOR HBV
Serum Specim en (log10Titer)
05382874001-10EN
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Figure 20
and the VERSANT HBV DNA 3.0 Assay

8.0
7.0
6.0
5.0
4.0
3.0
2.0
Y = 0.892 * X + 0.260
R 2 = 0.810
1.0
0.0
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
VERSANT HBV DNA 3.0 As s ay

05382874001-10EN
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Figure 21
and the VERSANT HBV DNA 3.0 Assay

Serum Specimen (Log10 Titer)
Y = 0.913 * X + 0.125
R 2 = 0.837
0
0
VERSANT HBV DNA 3.0 Assay

Serum Specim en (log Titer)
10
05382874001-10EN
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05382874001-10EN
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08/2013
Remove All rights reserved from copyright statement.

Correct Technical Support Telephone number from 1-800-428-2336 to 1-800-526-1247.
Please contact your local Roche Representative if you have any questions.
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U.S. patents #5,677,124, #5,919,625, #5,939,262, #6,214,982, #6,399,307, and other patents pending.
ARMORED RNA is a trademark of Ambion and Cenetron.
MICROSOFT and WINDOWS are trademarks of Microsoft Corporation in the United States and/or other
countries.
PROCLIN is a trademark of Rohm and Haas Company.
SST is a trademark of Becton Dickinson and Co.
VERSANT is a trademark of Bayer Corporation.
The cyanine dyes in this product are subject to patent rights of GE Healthcare Bio-Sciences Corp. and
Carnegie Mellon University and are licensed to Roche solely for incorporation into components of research
and in-vitro diagnostic kits. Any use of such kits for purposes other than research or in-vitro diagnostics
requires sublicenses from GE Healthcare Bio-Sciences Corp., Piscataway, New Jersey, U.S.A. and Carnegie
Mellon University, Pittsburgh, Pennsylvania, U.S.A.
Certain components of this product are covered by one or more US Patents and their foreign equivalents
issued to Isis Pharmaceuticals and licensed to Roche Molecular Systems, Inc.
Carryover prevention technology is covered by US Patent 5,035,996 and foreign counterpart patents owned by
Life Technologies, Inc. and is licensed to Roche Molecular Systems, Inc.
2013 Roche Molecular Systems, Inc.
08/2013
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05382874001-10EN
05382874001-10
43
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The following symbols are now used in labeling for Roche PCR diagnostic products.
Ancillary Software
In Vitro Diagnostic Medical Device
Authorised Representative
in the European Community
Lower Limit of Assigned Range
Barcode Data Sheet
Manufacturer
Batch code
Store in the dark
Biological Risks
Sufficient For
Catalogue Number
Temperature Limitation
SW
Consult instructions for use
Test Definition File

TDF
Contents of kit
Upper Limit of Assigned Range
Distributed by
Use By
For IVD Performance Evaluation Only
This product fulfills the requirements of the European Directive 98/79 EC for
in vitro diagnostic medical devices.
US Customer Technical Support 1-800-526-1247
05382874001-10EN
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COBAS AmpliPrep/COBAS TaqMan

HBV Test, version 2.0

P/N: 04894570 190

COBAS AmpliPrep/COBAS TaqMan

P/N: 03587797 190

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Detection of PCR Products in a COBAS TaqMan Test

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HBV DNA Quantitation

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93% (w/w) Isopropanol

< 5% (w/w) Polydocanol

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HBV QS, v2.0

HBV MMX, v2.0

HBV H(+)C, v2.0

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HBV L(+)C, v2.0

0.1% ProClin 300 preservative

HBV L(+)C, v2.0 Clip

COBAS AmpliPrep/COBAS TaqMan Wash Reagent

WARNINGS AND PRECAUTIONS

FOR IN VITRO DIAGNOSTIC USE.

Do not pipette by mouth.

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Do not mix reagent cassettes or controls from different kits.

Do not use a kit after its expiration date.

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STORAGE AND HANDLING REQUIREMENTS

Do not freeze reagents or controls.

COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0

COBAS AmpliPrep/COBAS TaqMan Wash Reagent

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MATERIALS REQUIRED BUT NOT PROVIDED

COBAS AmpliPrep Instrument

AMPLILINK Software v3.2 or v3.3 Series Manuals:

Sample processing units: SPUs

OTHER MATERIALS REQUIRED BUT NOT PROVIDED

Sample Rack (SK 24 rack)

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SPECIMEN COLLECTION, TRANSPORT AND STORAGE

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INSTRUCTIONS FOR USE

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COBAS AmpliPrep Instrument Set-up

Maintenance and Priming

The COBAS AmpliPrep Instrument is ready for operation in stand-by mode.

Log onto the Windows XP operating system.

Double click the AMPLILINK software icon.

Loading of Reagent Cassettes

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COBAS TaqMan Analyzer or COBAS TaqMan 48 Analyzer

Racks, Carriers and

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Ordering and Loading of Specimens

Print the Sample Rack Order report to use as a worksheet.

Start of COBAS AmpliPrep Instrument Run

Check for flags or error messages.

Remove waste from the COBAS AmpliPrep Instrument.