en

HBsAg Qualitative II
system
2G22
G2-3097/R03
B2G220
Read Highlighted Changes
Revised November 2011

HBsAg Qualitative II
Customer Service: Contact your local representative or find country specific contact information
on www.abbottdiagnostics.com
Package insert instructions must be carefully followed. Reliability of assay results cannot be guaranteed if there are any
deviations from the instructions in this package insert.

Key to symbols used

List Number Serial Number
In Vitro Diagnostic Medical
Device Control Number
Lot Number
Reaction Vessels
Expiration Date
Reagent Lot
Store at 2-8°C
Replacement Caps

Caution
Sample Cups

Consult instructions for use Septum

Warning: May cause an

Manufacturer allergic reaction

See REAGENTS section for a full explanation of symbols used in reagent

component naming.

1
NAME Other Reagents
ARCHITECT HBsAg Qualitative II ARCHITECT i Pre-Trigger Solution
INTENDED USE • Pre-trigger solution containing 1.32% (w/v)
hydrogen peroxide.
The ARCHITECT HBsAg Qualitative II assay is a chemiluminescent
microparticle immunoassay (CMIA) for the qualitative detection of ARCHITECT i Trigger Solution
hepatitis B surface antigen (HBsAg) in human serum and plasma. • Trigger solution containing 0.35 N sodium
The ARCHITECT HBsAg Qualitative II assay is intended to be used as an hydroxide.
aid in the diagnosis of HBV infection and as a screening test for donated ARCHITECT i Wash Buffer
blood and plasma. • Wash buffer containing phosphate buffered saline
solution. Preservatives: antimicrobial agents.
SUMMARY AND EXPLANATION OF TEST
The causative agent of serum hepatitis is hepatitis B virus (HBV) which WARNINGS AND PRECAUTIONS
is an enveloped DNA virus. During infection, HBV produces an excess of •
hepatitis B surface antigen (HBsAg), also known as Australia antigen, which • For In Vitro Diagnostic Use
can be detected in the blood of infected individuals. It is responsible for • Package insert instructions must be carefully followed. Reliability of
binding the virus to the liver cell and is the target structure of neutralizing assay results cannot be guaranteed if there are any deviations from
antibodies.1,2 HBsAg is the first serological marker after infection with the instructions in this package insert.
HBV appearing one to ten weeks after exposure and two to eight weeks
before the onset of clinical symptoms.3,4 HBsAg persists during this acute
Safety Precautions
phase and clears late in the convalescence period. Failure to clear HBsAg
within six months indicates a chronic HBsAg carrier state.
• CAUTION: This product contains human sourced and/or
HBsAg assays are used to identify persons infected with HBV and to potentially infectious components. Refer to the REAGENTS section
prevent transmission of the virus by blood and blood products as well of this package insert. No known test method can offer complete
as to monitor the status of infected individuals in combination with other assurance that products derived from human sources or inactivated
hepatitis B serological markers.5 In most countries, testing for HBsAg is microorganisms will not transmit infection. Therefore, all human
part of the antenatal screening program to identify HBV infected mothers sourced materials should be considered potentially infectious. It is
and to prevent perinatal HBV infection by subsequent immunization.6 recommended that these reagents and human specimens be handled
BIOLOGICAL PRINCIPLES OF THE PROCEDURE in accordance with the OSHA Standard on Bloodborne Pathogens7.
The ARCHITECT HBsAg Qualitative II assay is a one-step immunoassay Biosafety Level 28 or other appropriate biosafety practices9,10 should
for the qualitative detection of HBsAg in human serum and plasma using be used for materials that contain or are suspected of containing
CMIA technology, with flexible assay protocols, referred to as Chemiflex. infectious agents.
(Note: Ancillary Wash Buffer is added in a second incubation step, so the • The Conjugate contains human plasma that is nonreactive for HBsAg,
assay file performs a two-step assay protocol). HIV-1 Ag or HIV-1 RNA, anti-HIV-1/HIV-2, and anti-HCV.
In the ARCHITECT HBsAg Qualitative II assay, sample, anti-HBs coated • The following warnings and precautions apply to these components:
paramagnetic microparticles, and anti-HBs acridinium-labeled conjugate •• Microparticles
are combined to create a reaction mixture. HBsAg present in the •• Conjugate
sample binds to the anti-HBs coated microparticles and to the anti‑HBs •• Ancillary Wash Buffer
acridinium‑labeled conjugate. After washing, ancillary wash buffer is
added to the reaction mixture. Following another wash cycle, pre-trigger
and trigger solutions are added to the reaction mixture. The resulting
chemiluminescent reaction is measured as relative light units (RLUs). A
direct relationship exists between the amount of HBsAg in the sample and WARNING: Contains methylisothiazolones
the RLUs detected by the ARCHITECT i System optics. H317 May cause an allergic skin reaction.
The presence or absence of HBsAg in the sample is determined by Prevention
comparing the chemiluminescent signal in the reaction to the cutoff signal P261 Avoid breathing mist / vapours / spray.
determined from an active calibration. If the chemiluminescent signal in P272 Contaminated work clothing should not be allowed
the specimen is greater than or equal to the cutoff signal, the sample is out of the workplace.
considered reactive for HBsAg. P280 Wear protective gloves / protective
For additional information on system and assay technology, refer to the clothing / eye protection.
ARCHITECT System Operations Manual, Section 3. Response
REAGENTS P302+P352 IF ON SKIN: Wash with plenty of water.
Reagent Kit, 100 Tests/500 Tests P333+P313 If skin irritation or rash occurs: Get medical
NOTE: Some kit sizes are not available in all countries or for use on all advice / attention.
ARCHITECT i Systems. Please contact your local distributor. P363 Wash contaminated clothing before reuse.
ARCHITECT HBsAg Qualitative II Reagent Kit (2G22) This material and its container must be disposed of in a safe way.
• 1 or 4 bottles (6.6 mL per 100-test bottle / 27.0 mL • For a detailed discussion of safety precautions during system operation,
per 500-test bottle) anti-HBs (mouse, monoclonal, IgM, IgG) coated refer to the ARCHITECT System Operations Manual, Section 8.
microparticles in MES buffer with protein (bovine serum albumin) Handling Precautions
stabilizer. Minimum concentration: 0.08% solids. Preservatives: • Do not use reagents kits beyond the expiration date.
ProClin 300 and ProClin 950.
• Do not pool reagents within a kit or between reagent kits.
• 1 or 4 bottles (5.9 mL per 100-test bottle/26.3 mL per
• Before loading the ARCHITECT HBsAg Qualitative II Reagent Kit on
500-test bottle) anti-HBs (mouse, monoclonal, IgG) and anti-HBs
the system for the first time, the microparticle bottle requires mixing
(goat, IgG) acridinium-labeled conjugate in phosphate buffer with
to resuspend microparticles that may have settled during shipment.
human plasma and protein (bovine serum albumin, fetal bovine serum,
For microparticle mixing instructions, refer to the PROCEDURE, Assay
goat IgG, mouse IgG) stabilizers. Minimum concentration: 0.35 μg/mL.
Procedure section of this package insert.
Preservatives: ProClin 300 and ProClin 950.
• Septums MUST be used to prevent reagent evaporation and
• 1 or 4 bottles (5.9 mL per 100-test bottle/
contamination and to ensure reagent integrity. Reliability of assay
26.3 mL per 500-test bottle) ancillary wash buffer containing MES
results cannot be guaranteed if septums are not used according to
buffer. Preservatives: ProClin 300 and ProClin 950.
the instructions in this package insert.

2
• To avoid contamination, wear clean gloves when placing a septum on • Performance has not been established for the use of body fluids other
an uncapped reagent bottle. than human serum or plasma or for cadaveric specimens.
•• When handling conjugate vials, change gloves that have contacted • Liquid anticoagulants may have a dilution effect resulting in lower
human serum or plasma, since introduction of human IgG or IgM concentrations for individual patient specimens.
will result in a neutralized conjugate. • The ARCHITECT i System does not provide the capability to verify
•• Once a septum has been placed on the reagent bottle, do not specimen type. It is the responsibility of the operator to verify that
invert the bottle as this will result in reagent leakage and may the correct specimen types are used in the ARCHITECT HBsAg
compromise assay results. Qualitative II assay.
•• Over time, residual liquids may dry on the septum surface. These Specimen Conditions
are typically dried salts, and have no effect on assay efficacy. • Do not use specimens with the following conditions:
• For a detailed discussion of handling precautions during system •• heat-inactivated
operation, refer to the ARCHITECT System Operations Manual,
•• pooled
Section 7.
•• grossly hemolyzed
Storage Instructions •• obvious microbial contamination
• For accurate results, serum and plasma specimens should be free of
• The ARCHITECT HBsAg Qualitative II Reagent Kit must be fibrin, red blood cells, and other particulate matter. Serum specimens
stored at 2-8°C in an upright position and may be used immediately from patients receiving anticoagulant or thrombolytic therapy may
after removal from 2-8°C storage. contain fibrin due to incomplete clot formation.
• When stored and handled as directed, the reagents are stable until • Ensure that complete clot formation in serum specimens has taken
the expiration date. place prior to centrifugation. If the specimen is centrifuged before
• The ARCHITECT HBsAg Qualitative II Reagent Kit may be stored on a complete clot forms, the presence of fibrin may cause erroneous
board the ARCHITECT i System for a maximum of 30 days. After results.
30 days, the reagent kit must be discarded. For information on tracking • As specimens from heparinized patients may be partially coagulated
onboard time, refer to the ARCHITECT System Operations Manual, and erroneous results could occur due to the presence of fibrin, draw
Section 5. the specimen prior to heparin therapy.
• Reagents may be stored on or off the ARCHITECT i System. If reagents • Use caution when handling patient specimens to prevent cross
are removed from the system, store them at 2-8°C (with septums and contamination. Use of disposable pipettes or pipette tips is
replacement caps) in an upright position. For reagents stored off the recommended.
system, it is recommended that they be stored in their original trays and • For optimal results, inspect all specimens for bubbles. Remove bubbles
boxes to ensure they remain upright. If the microparticle bottle does with an applicator stick before analysis. Use a new applicator stick for
not remain upright (with a septum installed) while in refrigerated each specimen to prevent cross contamination.
storage off the system, the reagent kit must be discarded. For • No qualitative performance differences were observed between
information on unloading reagents, refer to the ARCHITECT System experimental controls and nonreactive or spiked reactive specimens
Operations Manual, Section 5. tested with elevated levels of conjugated or unconjugated bilirubin
Indications of Reagent Deterioration (20 mg/dL), triglycerides (3000 mg/dL), protein (12 g/dL), or
When a control value is out of the specified range, it may indicate hemoglobin (500 mg/dL).
deterioration of the reagents or errors in technique. Associated test Preparation for Analysis
results are invalid and samples must be retested. Assay recalibration may • Follow the tube manufacturer’s processing instructions for serum
be necessary. For troubleshooting information, refer to the ARCHITECT and plasma collection tubes. Gravity separation is not sufficient for
System Operations Manual, Section 10. specimen preparation.
INSTRUMENT PROCEDURE • Prepare frozen specimens as follows:
• The ARCHITECT HBsAg Qualitative II assay file (assay number 628) •• Frozen specimens must be completely thawed before mixing.
must be installed on the ARCHITECT i System before performing the •• Mix thawed specimens thoroughly by inverting 10 times or by
assay. low speed vortexing. Visually inspect the specimens. If layering
• For detailed information on assay file installation and viewing and or stratification is observed, continue mixing until specimens
editing assay parameters, refer to the ARCHITECT System Operations are visibly homogeneous. If samples are not mixed thoroughly,
Manual, Section 2. inconsistent results may be obtained.
• For information on printing assay parameters, refer to the ARCHITECT •• Centrifuge mixed specimens as described below.
System Operations Manual, Section 5. • To ensure consistency in results, specimens must be transferred to a
• For a detailed description of system procedures, refer to the centrifuge tube and centrifuged at >10,000 RCF (Relative Centrifugal
ARCHITECT System Operations Manual. Force) for 10 minutes before testing if
•• they contain fibrin, red blood cells, or other particulate matter or
SPECIMEN COLLECTION AND PREPARATION FOR ANALYSIS
•• they were frozen and thawed.
Specimen Types
• Centrifuged specimens with a lipid layer on the top must be transferred
• The specimen collection tubes listed below were verified for use with to a sample cup or secondary tube. Care must be taken to transfer
the ARCHITECT HBsAg Qualitative II assay. Other specimen collection only the clarified specimen without the lipemic material.
tubes have not been tested with this assay.
• Transfer clarified specimen to a sample cup or secondary tube for
•• Human serum (including serum collected in serum separator testing.
tubes)
•• Human plasma collected in: Storage
• Specimens may be stored on or off the clot, red blood cells, or
•• Lithium heparin •• Sodium heparin separator gel for
•• Dipotassium EDTA •• CPD •• up to 24 hours at room temperature or
•• Tripotassium EDTA •• CPDA-1 •• up to 6 days at 2-8°C.
•• Sodium citrate •• ACD • If testing will be delayed more than 6 days, remove serum or plasma
•• Plasma separator tubes •• Potassium oxalate / from the clot, red blood cells, or separator gel and store at - 20°C or
(lithium heparin) sodium fluoride plasma colder.
• Avoid more than 3 freeze/thaw cycles.

3
Shipping •• ≤ 3 hours on-board: 150 µL for the first HBsAg Qualitative II test
• Before shipping specimens, it is recommended that specimens be plus 75 µL for each additional HBsAg Qualitative II test from the
removed from the clot, red blood cells, or separator gel. same sample cup.
• When shipping specimens, package and label specimens in •• > 3 hours on-board: replace with a fresh sample (patient
compliance with applicable state, federal, and international regulations specimens, controls, and calibrators).
covering the transport of clinical specimens and infectious substances. •• If using primary or aliquot tubes, use the sample gauge to ensure
• Specimens may be shipped ambient, at 2-8°C (wet ice), or frozen sufficient patient specimen is present.
(dry ice). Do not exceed the storage time limitations listed above. • Prepare calibrators and controls.
PROCEDURE •• Mix the ARCHITECT HBsAg Qualitative II Calibrators and Controls
Materials Provided by gentle inversion before use.
• 2G22 ARCHITECT HBsAg Qualitative II Reagent Kit •• To obtain the recommended volume requirements for the
ARCHITECT HBsAg Qualitative II Calibrators and Controls, hold the
Materials Required but not Provided bottles vertically, and dispense 11 drops of each calibrator and
• ARCHITECT i System 6 drops of each control into each respective sample cup.
• ARCHITECT HBsAg Qualitative II Assay file, obtained from the •• If commercially available control material is used, follow the
•• ARCHITECT i System e-Assay CD-ROM obtained from manufacturer’s instructions for preparation.
www.abbottdiagnostics.com or • Load samples
•• ARCHITECT i System Assay CD-ROM •• For information on loading samples, refer to the ARCHITECT
• 2G22-01 ARCHITECT HBsAg Qualitative II Calibrators System Operations Manual, Section 5.
• 2G22-10 ARCHITECT HBsAg Qualitative II Controls or other control • Press RUN.
material • For additional information on principles of operation, refer to the
• ARCHITECT i ARCHITECT System Operations Manual, Section 3.
• ARCHITECT i • For optimal performance, it is important to perform routine maintenance
• ARCHITECT i as described in the ARCHITECT System Operations Manual, Section 9.
• ARCHITECT i Perform maintenance more frequently when required by laboratory
• ARCHITECT i procedures.
• ARCHITECT i Specimen Dilution Procedure
• ARCHITECT i Specimens cannot be diluted for the ARCHITECT HBsAg Qualitative II assay.
• Pipettes or pipette tips (optional) to deliver the specified volumes. Calibration
For information on materials required for maintenance procedures, refer to • To perform an ARCHITECT HBsAg Qualitative II calibration, test
the ARCHITECT System Operations Manual, Section 9. calibrators 1 and 2 in replicates of 3. The calibrators should be priority
Assay Procedure loaded.
• Before loading the ARCHITECT HBsAg Qualitative II Reagent Kit on • A single sample of each control level must be tested to evaluate the
the system for the first time, the microparticle bottle requires mixing assay calibration.
to resuspend microparticles that may have settled during shipment. •• Order controls as described in the Assay Procedure section.
After the first time the microparticles have been loaded, no further •• Ensure that assay control values are within the ranges specified in
mixing is required. the control package insert.
•• Invert the microparticle bottle 30 times. • Once an ARCHITECT HBsAg Qualitative II calibration is accepted
•• Visually inspect the bottle to ensure microparticles are and stored, all subsequent samples may be tested without further
resuspended. If microparticles are still adhered to the bottle, calibration unless:
continue to invert the bottle until the microparticles have been •• A reagent kit with a new lot number is used.
completely resuspended. •• Controls are out of range.
•• If the microparticles do not resuspend, DO NOT USE. Contact • For detailed information on how to perform an assay calibration, refer
your Abbott representative. to the ARCHITECT System Operations Manual, Section 6.
•• Once the microparticles have been resuspended, place a septum
QUALITY CONTROL PROCEDURES
on the bottle. For instructions about placing septums on bottles,
The recommended control requirement for the ARCHITECT HBsAg
refer to the Handling Precautions section of this package insert.
Qualitative II assay is that a single sample of each control be tested
• Load the ARCHITECT HBsAg Qualitative II Reagent Kit on the once every 24 hours each day of use. If your laboratory quality control
ARCHITECT i System. procedures require more frequent use of controls to verify test results,
•• Verify that all necessary reagents are present. follow those procedures. Additional controls may be tested in conformance
•• Ensure that septums are present on all reagent bottles. with local, state, and/or federal regulations or accreditation requirements
• Order calibration, if necessary. and your laboratory’s quality control policy.
•• For information on ordering calibrations, refer to the ARCHITECT Control values must be within the ranges specified in the control package
System Operations Manual, Section 6. insert. If a control result is out of its specified range, any test results
• Order tests. generated since the last acceptable control results must be evaluated
•• For information on ordering patient specimens and controls and to determine if test results may have been adversely affected. Adversely
for general operating procedures, refer to the ARCHITECT System affected test results are invalid, and these samples must be retested. For
Operations Manual, Section 5. troubleshooting information, refer to the ARCHITECT System Operations
• The minimum sample cup volume is calculated by the system and Manual, Section 10.
is printed on the Orderlist report. No more than 10 replicates may Verification of Assay Claims
be sampled from the same sample cup. To minimize the effects of For protocols to verify package insert claims, refer to the ARCHITECT
evaporation, verify adequate sample cup volume is present before System Operations Manual, Appendix B. The ARCHITECT HBsAg
running the test. Qualitative II assay belongs to method group 5, except functional
•• Priority: 125 µL for the first HBsAg Qualitative II test plus 75 µL for sensitivity.
each additional HBsAg Qualitative II test from the same sample
cup.

4
RESULTS Precision
Calculations The ARCHITECT HBsAg Qualitative II assay is designed to have an
• The ARCHITECT i System calculates the result for the ARCHITECT imprecision of ≤ 10% within-laboratory (Total) CV for the positive control
HBsAg Qualitative II assay using the ratio of the sample RLU to the and low positive panel and a standard deviation (SD) of ≤ 0.10 S/CO for
cutoff RLU (S/CO) for each specimen and control. the high negative panel.
•• Cutoff RLU = (0.0575 x Calibrator 1 Mean RLU) + Within-Laboratory Precision
(0.8 x Calibrator 2 Mean RLU) A study was performed based on guidance from the National Committee
•• S/CO = Sample RLU/Cutoff RLU for Clinical Laboratory Standards (NCCLS) document EP5-A2.14 Testing
was conducted using 3 lots of ARCHITECT HBsAg Qualitative II reagents,
Interpretation of Results calibrators, and controls, and 4 instruments. Two controls and two panels
ARCHITECT HBsAg Qualitative II Initial Result were assayed in a minimum of 2 replicates at 2 separate times per day
Instrument for 20 different days. Each reagent lot used a single calibration curve
Initial Result (S/CO) Interpretation Retest Procedure throughout the study. The ranges for all instruments and reagent lots are
< 1.00 NONREACTIVE No retest required. summarized across instruments and reagent lots in the following table.
≥ 1.00 REACTIVE Retest in duplicate. Within-Laboratory
Within-Run Precision (Total)
• Initially reactive specimens require retesting. Specimens that contain Mean
Range Range
particulate matter should be recentrifuged according to directions in Range
the SPECIMEN COLLECTION AND PREPARATION FOR ANALYSIS Sample n S/CO SD %CV SD %CV
section in this package insert. Negative
956 0.15 - 0.18 0.012 - 0.016 NA 0.014 - 0.030 NA
Control
ARCHITECT HBsAg Qualitative II Retest Results
Positive
Instrument 958 3.26 - 3.45 0.056 - 0.082 1.7 - 2.5 0.072 - 0.103 2.1 - 3.2
Control
Interpretation Specimen Classification
High
Both results nonreactive Specimen considered negative for HBsAg. Negative 955 0.71 - 0.77 0.021 - 0.024 NA 0.025 - 0.033 NA
One or both results Specimen considered repeatedly reactive; Panel
reactive confirm using a neutralizing assay.* Low
* The ARCHITECT HBsAg Qualitative II Confirmatory assay is recommended. Positive 956 1.17 - 1.27 0.026 - 0.040 2.1 - 3.4 0.029 - 0.048 2.3 - 4.1
Panel
• Confirm repeatedly reactive specimens using a neutralizing assay
(e.g., ARCHITECT HBsAg Qualitative II Confirmatory) before disclosing NA = Not applicable
HBsAg status to the patient. System Reproducibility
For details on configuring the ARCHITECT i System to use gray zone A 5-day precision study was performed for the ARCHITECT HBsAg
interpretations, refer to the ARCHITECT System Operations Manual, Qualitative II assay based on guidance from the Clinical and Laboratory
Section 2. The grayzone interpretations are editable parameters, and Standards Institute (CLSI) document EP15-A2.15 Testing was conducted
should be utilized per end user requirements. at 3 clinical sites using 3 lots each of ARCHITECT HBsAg Qualitative II
Flags reagents, calibrators, and controls per site. Two controls and 2 panels
Some results may contain information in the Flags field. For a description were assayed in replicates of 4 at 2 separate times of day for 5 days. The
of the flags that may appear in this field, refer to the ARCHITECT System data are summarized in the following table.
Operations Manual, Section 5. Within-
LIMITATIONS OF THE PROCEDURE Laboratory
Grand Precision
• If the ARCHITECT HBsAg Qualitative II results are inconsistent with
Mean Within-Run Within-Day (Total)
clinical evidence, additional testing is suggested to confirm the result.
• For diagnostic purposes, results should be used in conjunction with Sample n S/CO SD %CV SD %CV SD %CV
patient history and other hepatitis markers for diagnosis of acute and Negative
360 0.17 0.028 NA 0.031 NA 0.031 NA
chronic infection. Control
• Specimens from patients who have received preparations of mouse Positive
360 3.45 0.066 1.9 0.070 2.0 0.073 2.1
monoclonal antibodies for diagnosis or therapy may contain human Control
anti-mouse antibodies (HAMA).11,12 Specimens containing HAMA High Negative
may produce anomalous values when tested with assay kits such 360 0.77 0.037 4.8 0.061 7.9 0.061 7.9
Panel
as ARCHITECT HBsAg Qualitative II that employ mouse monoclonal Low Positive
antibodies.11 360 1.28 0.066 5.1 0.066 5.1 0.066 5.1
Panel
• Heterophilic antibodies in human serum can react with reagent
immunoglobulins, interfering with in vitro immunoassays.13 Patients NA = not applicable
routinely exposed to animals or to animal serum products can be Specificity
prone to this interference and anomalous results may be observed. Blood Donor Specimens
Additional information may be required for diagnosis. The ARCHITECT HBsAg Qualitative II assay is designed to have a
• Refer to the SPECIMEN COLLECTION AND PREPARATION FOR specificity of > 99.5% on blood donor specimens.
ANALYSIS section of this package insert for specimen limitations. A study was performed at three external sites on a total of 5401 serum
SPECIFIC PERFORMANCE CHARACTERISTICS and plasma specimens collected from two blood-donation centers. For
All performance studies were conducted using the ARCHITECT 1 specimen which was tested as initial and repeat reactive on ARCHITECT
i2000 / i2000SR Systems. Additionally, the Within-Laboratory Precision, HBsAg Qualitative II, the presence of HBsAg was confirmed by specific
Analytical Sensitivity, and Seroconversion studies were conducted using neutralization with anti-HBs. The specificity on the remaining 5400 blood
the ARCHITECT i1000SR. donors was assessed to be 99.91% (5395/5400) with an assumed zero
Assay results obtained in individual laboratories may vary from data prevalence of HBV infection. The data are summarized in the following
presented. table.

5
 IRa RRa 95% Confidence Analytical Specificity
Category n (%) (%) Specificity Interval The ARCHITECT HBsAg Qualitative II assay was evaluated for potential
cross-reactivity for specimens from individuals with medical conditions
Overall
7 6 99.91% unrelated to HBV infection. A total of 294 specimens from 28 different
Blood 5401b 99.78% - 99.97%
(0.13%) (0.11%) (5395/5400) categories were tested. Two hundred ninety specimens were nonreactive
Donors
and 4 specimens were reactive by the ARCHITECT HBsAg Qualitative II
Blood and commercially available HBsAg assays. All 4 reactive specimens were
4 3 99.89%
Donors 2700 99.68% - 99.98% confirmed positive for HBsAg by the ARCHITECT HBsAg Qualitative II
(0.15%) (0.11%) (2697/2700)
Plasma Confirmatory and commercially available HBsAg confirmatory assays. The
Blood data are summarized by final interpretation in the following table.
3 3 99.93%
Donors 2701b 99.73% - 99.99%
(0.11%) (0.11%) (2698/2700) Commercially Available
Serum
HBsAg Assay
a IR = Initially Reactive, RR = Repeatedly Reactive Nonreactive Reactive
b One specimen confirmed positive. ARCHITECT ARCHITECT
Diagnostic Specimens HBsAg HBsAg
A study was performed using a total of 1499 randomly selected diagnostic Qualitative II Qualitative II
patients, including specimens from hospitalized and hemodialysis patients. Category n NRa Ra NRa Ra
For 16 specimens which were tested as initial and repeat reactives on Cytomegalovirus (CMV) 10 10 0 0 0
ARCHITECT HBsAg Qualitative II, the presence of HBsAg was confirmed
Epstein-Barr Virus (EBV) 10 10 0 0 0
by specific neutralization with anti-HBs. The specificity on the remaining
1483 diagnostic specimens was assessed to be 99.93% (1482/1483) with Multiple Transfusion Recipients 10 10 0 0 0
an assumed zero prevalence of HBV infection. The data are summarized Hepatitis A Virus (HAV) 10 10 0 0 0
in the following table. Human Anti-Mouse
15 15 0 0 0
IRa RRa 95% Confidence Antibodies (HAMA) Positive
Category n (%) (%) Specificityb Interval Hepatitis C Virus (HCV) 10 10 0 0 0
Overall 18 17 99.93% Human Immunodeficiency Virus
1499c 99.62% - 100.00% 10 10 0 0 0
Diagnostics (1.20%) (1.13%) (1482/1483) (HIV-1)
Hospitalized/ 12 11 99.90% Autoimmune Hepatitis 10 10 0 0 0
999d 99.44% - 100.00%
Diagnostics (1.20%) (1.10%) (988/989) Human Immunodeficiency Virus
17 14 0 0 3
6 6 100.00% (HIV-2)
Hemodialysis 500e 99.26% - 100.00%
(1.20%) (1.20%) (494/494) Fatty Liver Disease 10 10 0 0 0
a Herpes Simplex Virus (HSV) 10 10 0 0 0
IR = Initially Reactive, RR = Repeatedly Reactive
b Hepatocellular Carcinoma 10 10 0 0 0
One aberrant result was observed and the specificity was 99.93%
(1481/1482) with this specimen excluded. Human T-Lymphotropic Virus
9 9 0 0 0
c Sixteen specimens confirmed positive. (HTLV-1/2)
d Ten specimens confirmed positive. T. pallidum 2 2 0 0 0
e Six specimens confirmed positive. N. gonorrhea 9 9 0 0 0
C. trachomatis 7 7 0 0 0
Sensitivity
The ARCHITECT HBsAg Qualitative II assay is designed to show sensitivity T. cruzi 10 10 0 0 0
performance that is greater than or equal to the lower limit of the 95% Rheumatoid Factor (RF) 10 10 0 0 0
confidence interval for a commercially available HBsAg assay on the Anti-Nuclear Antibodies (ANA) 10 10 0 0 0
same population of HBsAg positive specimens. Pregnancy 1st Trimester 15 15 0 0 0
For the 402 HBsAg positive specimens from patients with unknown disease Pregnancy 2nd Trimester 15 14 0 0 1
status evaluated in this study, the lower limit of the 95% confidence
Pregnancy 3rd Trimester 15 15 0 0 0
interval for the commercially available HBsAg assay was 99.09%. In this
study, the sensitivity of the ARCHITECT HBsAg Qualitative II assay was Multiparous Females 10 10 0 0 0
100.00% (402/402). IgM Monoclonal Gammopathy 10 10 0 0 0
Analytical Sensitivity IgG Monoclonal Gammopathy 10 10 0 0 0
The ARCHITECT HBsAg Qualitative II assay is designed to have a mean Multiple Myeloma 10 10 0 0 0
analytical sensitivity value that is less than or equal to the lower limit Influenza Vaccine Recipients 10 10 0 0 0
of the 95% confidence interval around the mean analytical sensitivity Hemodialysis Patient 10 10 0 0 0
of a commercially available HBsAg assay. Analytical sensitivity was Total 294 290 0 0 4
evaluated using serial dilutions of the WHO 2nd International HBsAg
a NR = Nonreactive, R = Reactive
Standard (subtype adw2, genotype A, NIBSC Code 00/588). The dilutions
ranged from 0.010 to 0.5 IU/mL. Recalcified negative human plasma/ Seroconversion Sensitivity
serum was used as the diluent and represented the 0 IU/mL sample. The ARCHITECT HBsAg Qualitative II assay is designed to have a
The dilutions were tested across 3 reagent lots on 3 instrument types seroconversion sensitivity that is better than or equivalent to the
(1 i2000SR, 1 i2000, and 1 i1000SR). In this study, the lower limit of the seroconversion sensitivity of a commercially available HBsAg assay. To
95% confidence interval for the commercially available HBsAg assay determine the seroconversion sensitivity, 30 HBV seroconversion panels
was 0.021 IU/mL. The analytical sensitivity results for ARCHITECT obtained from commercial vendors were tested using the ARCHITECT
HBsAg Qualitative II, calculated by linear regression, ranged from 0.017 HBsAg Qualitative II and ARCHITECT HBsAg Qualitative II Confirmatory
to 0.022 IU/mL. The mean analytical sensitivity ranged from 0.019 to assays. The results were compared to a commercially available HBsAg
0.020 IU/mL across instrument types. assay and representative data from 6 panels are summarized in the
following table.

6
 ARCHITECT HBsAg Commercially Available Final Interpretation
Days since Qualitative II S/CO HBsAg Assay S/CO ARCHITECT HBsAg Commercially Available
Panel ID 1st bleed Reactive ≥ 1.00 S/CO Reactive ≥ 1.00 S/CO Mutant Qualitative II HBsAg
0 0.31 0.39 Gln-129-His Repeatedly Reactive Repeatedly Reactive
3 0.74 0.70 Met-133-Leu Repeatedly Reactive Repeatedly Reactive
6271 7 1.88 1.81 Asp-144-Ala Repeatedly Reactive Nonreactive
12 14.41 9.49 Gly-145-Arg Repeatedly Reactive Repeatedly Reactive
18 113.86 56.70 Thr-123-Ala Repeatedly Reactive Nonreactive
0 0.59 0.64 P142L+G145R Repeatedly Reactive Repeatedly Reactive
4 1.32 0.91 P142S+G145R Repeatedly Reactive Repeatedly Reactive
PHM 925 8 2.48 1.87 122NT Repeatedly Reactive Repeatedly Reactive
14 5.69 4.10 122RA Repeatedly Reactive Repeatedly Reactive
17 6.72 3.46
HBV Genotype Detection
0 0.50 0.41 The ARCHITECT HBsAg Qualitative II assay is designed to detect HBV
3 4.95 2.28 genotypes A through F and H. A study was performed to evaluate the
PHM 930 8 43.38 19.73 ability of the ARCHITECT HBsAg Qualitative II assay to detect different
12 124.59 47.42 HBV genotypes by testing a commercially available genotype panel
containing genotypes A through F and H. A total of 18 panel members
15 321.30 112.32 (3 panel members each of A, B, C, D, and E; 2 panel members of F,
2 0.79 0.69 and 1 panel member of H) were tested using the ARCHITECT HBsAg
Qualitative II and ARCHITECT HBsAg Qualitative II Confirmatory assays.
7 4.01 2.26
All genotypes were reactive by the ARCHITECT HBsAg Qualitative II
PHM 933 9 9.07 4.85 assay and confirmed positive by the ARCHITECT HBsAg Qualitative II
16 45.03 22.30 Confirmatory assay.
144 2715.52 823.14 BIBLIOGRAPHY
0 0.17 0.72 1. Neurath AR, Kent SB, Strick N, et al. Identification and chemical
3 0.16 0.39 synthesis of a host cell receptor binding site on hepatitis B virus. Cell
1986;46:429-36.
7 0.25 0.55
6273 2. Szmuness W, Stevens CE, Harley EJ, et al. Hepatitis B vaccine-
14 1.05 1.02 demonstration of efficacy in a controlled clinical trial in a high-risk
25 20.99 13.84 population in the United States. N Engl J Med 1980;303:833-41.
30 158.83 73.40 3. Krugman S, Giles JP. Viral hepatitis, type B (MS-2-Strain)- further
0 0.36 0.42 observations on natural history and prevention. N Engl J Med
1973;288:755-60.
2 0.49 0.60
4. Krugman S, Overby LR, Mushahwar IK, et al. Viral hepatitis,
7 1.59 1.55 type B‑studies on natural history and prevention re-examined. N Engl
11002
9 2.40 2.08 J Med 1979;300:101-6.
35 1612.66 379.66 5. Perrillo RP, Aach RD. The clinical course and chronic sequelae of
39 403.92 232.20 hepatitis B virus infection. Seminars in Liver Disease 1981;1:15-25.
6. CDC. A comprehensive immunization strategy to eliminate
HBsAg Mutant Detection transmission of Hepatitis B virus infection in the United States:
The hepatitis B virus, unlike other DNA viruses, replicates through reverse recommendations of the Advisory Committee on Immunization
transcription. The reverse transcription process lacks proofreading Practices (ACIP) Part 1: Immunization of Infants, Children, and
capability; therefore, HBV is subject to a mutation rate 10 times higher Adolescents. MMWR 2005;54(RR-16):1-23.
than the mutation rate of other DNA viruses.16 Some of these mutations 7. US Department of Labor, Occupational Safety and Health
may cause changes in the antigenic structure of HBsAg, resulting in Administration, 29 CFR Part 1910.1030, Bloodborne pathogens.
epitopes that are no longer recognized by anti-HBs. HBsAg mutants 8. US Department of Health and Human Services. Biosafety in
have been reported in a wide range of patient populations, including Microbiological and Biomedical Laboratories. 5th ed. Washington, DC:
blood donors, vaccine recipients, renal dialysis patients, orthotopic liver US Government Printing Office; January 2007.
transplant recipients, infants born to HBsAg-positive mothers, and patients 9. World Health Organization. Laboratory Biosafety Manual 3rd ed.
undergoing nucleoside analog treatment for HBV.16-23 HBsAg mutations Geneva: World Health Organization; 2004.
may result in a less favorable outcome in some patients16,17,19 and false
10. Clinical and Laboratory Standards Institute. Protection of Laboratory
negative results in some HBsAg assays.16-18
Workers from Occupationally Acquired Infections: Approved
The ARCHITECT HBsAg Qualitative II assay is designed to have the ability Guideline—Third Edition. CLSI Document M29-A3. Wayne, PA:
to better detect (as reactive) the HBsAg mutant Thr-123-Ala and to have Clinical and Laboratory Standards Institute; 2005.
the equivalent or better ability to detect (as reactive) other HBsAg mutants
11. Primus FJ, Kelley EA, Hansen HJ, et al. “Sandwich”-type immunoassay
when compared to the comparator assay. A panel of 9 recombinant HBsAg
of carcinoembryonic antigen in patients receiving murine monoclonal
mutant samples was obtained. Each panel member was diluted with
antibodies for diagnosis and therapy. Clin Chem 1988;34(2):261-4.
recalcified negative human plasma to an S/CO of 2.0 ± 0.5 and tested
with the ARCHITECT HBsAg Qualitative II assay and with a comparator 12. Schroff RW, Foon KA, Beatty SM, et al. Human anti-murine
assay. The data are summarized in the following table. immunoglobulin responses in patients receiving monoclonal antibody
therapy. Cancer Res 1985;45:879-85.
13. Boscato LM, Stuart MC. Heterophilic antibodies: a problem for all
immunoassays. Clin Chem 1988;34(1):27-33.
14. National Committee for Clinical Laboratory Standards (NCCLS).
Evaluation of Precision Performance of Quantitative Measurement
Methods; Approved Guideline–Second Edition. NCCLS Document
EP5-A2. Wayne, PA: NCCLS; 2004.

7
15. Clinical and Laboratory Standards Institute. User Verification of
Performance for Precision and Trueness; Approved Guideline -
Second Edition. CLSI Document EP15-A2. Wayne, PA: CLSI; 2005.
16. Hunt CM, McGill JM, Allen MI, et al. Clinical relevance of hepatitis B
viral mutations. Hepatology 2000;31(5):1037‑44.
17. Locarnini SA. Hepatitis B virus surface antigen and polymerase gene
variants: potential virological and clinical significance. Hepatology
1998;27(1):294–7.
18. Zuckerman AJ. Effect of hepatitis B virus mutants on efficacy of
vaccination. Lancet 2000;355:1382‑4.
19. Carman WF, Trautwein C, Van Deursen FJ, et al. Hepatitis B virus
envelope variation after transplantation with and without hepatitis B
immune globulin prophylaxis. Hepatology 1996;24(3):489‑93.
20. Grethe S, Monazahian M, Böhme I, et al. Characterization of unusual
escape variants of hepatitis B virus isolated from a hepatitis B surface
antigen-negative subject. J Virology 1998;72(9):7692‑6.
21. Nainan OV, Stevens CE, Taylor PE, et al. Hepatitis B virus (HBV)
antibody resistant mutants among mothers and infants with chronic
HBV infection. In: Rizzetto M, Purcell RH, Gerin JL, et al., eds. Viral
Hepatitis and Liver Disease. Minerva Medica: Torino;1997:132‑134.
22. Jongerius JM, Wester M, Cuypers HTM, et al. New hepatitis B virus
mutant form in a blood donor that is undetectable in several hepatitis B
surface antigen screening assays. Transfusion 1998;38:56‑9.
23. Bock CT, Tillmann HL, Torresi J, et al. Selection of hepatitis B virus
polymerase mutants with enhanced replication by lamivudine treatment
after liver transplantation. Gastroenterology 2002;122:264‑73.

The following U.S. Patents are relevant to the ARCHITECT System or its
components. There are other such patents and patent applications in the
United States and worldwide.
5 468 646 5 543 524 5 545 739
5 565 570 5 669 819 5 783 699

ARCHITECT and Chemiflex are trademarks of Abbott Laboratories in

various jurisdictions.
ProClin is property of its respective owner.

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