DiaSorin S.p.A.

Via Crescentino snc - 13040 Saluggia (VC) - Italy

www.diasorin.com
Tel. +39.0161.4871

Changes: §1, §7, §9, §14, §15.1, §15.6, References;

Deletions: -

LIAISON ® XL HBsAg Quant ([REF] 310250)

1. INTENDED USE
The LIAISON ® XL MUREX HBsAg Quant assay uses chemiluminescent immunoassay (CLIA) technology for the quantitative
determination of Hepatitis B surface Antigen (HBsAg) in human serum and plasma samples included specimens collected
post-mortem (non-heart beating).
The assay is intended as an aid in the diagnosis of HBV infection in individuals with or without symptoms of hepatitis and
in the monitoring of treatment and disease status. It is also intended as a screening test for blood and hemocomponents
donors as well as for organ, tissue and cells post-mortem donors.
The test has to be performed on the LIAISON ® XL Analyzer only.

2. SUMMARY AND EXPLANATION OF THE TEST

Hepatitis is an inflammatory disease of the liver that can severely damage the organ. The disease can result from
non-infectious causes or from infectious viral and bacterial agents (5).
Viral hepatitis B is endemic throughout the world (11, 13, 20). The infection is spread primarily through percutaneous contact
with infected blood, e.g., sharing of needles by drug addicts or transfusion of blood products that have not been screened
for HBV (2, 5, 11). The hepatitis B virus (HBV) is also found in virtually every type of human body fluid and has been known
to be spread through oral and genital contact (2, 5, 11, 22). HBV can be transmitted perinatally from mother to child (2).
The incubation period for hepatitis B averages 90 days (range: 40-180 days). Common symptoms include malaise, fever,
gastroenteritis, and icterus (6). HBV infection can lead to (a) icteric hepatitis; (b) subclinical anicteric hepatitis; (c) fulminant
hepatitis; (d) chronic active or persistent hepatitis. Approximately 70% of patients with acute hepatitis B experience
subclinical or anicteric hepatitis, whereas 30% develop icteric hepatitis. Fulminant hepatitis due to massive
immune-mediated lysis of infected hepatocytes is unusual, occurring in approximately 0.1-0.5% of patients (5, 7, 11, 12, 15,
18, 21). In patients who recover from acute HBV infection, normalization of serum alanine amino-transferase (ALT) levels
usually occurs within one to four months. Persistently increased serum ALT levels for more than six months indicate
progression to chronic hepatitis B. The rate of progression from acute to chronic hepatitis B is affected by the age at
infection. The rate is up to 90% for perinatally acquired infection, 20-50% for infections between the age of one and five
years, and less than 5% for adult-acquired infection.
The complete hepatitis B virus (HBV) is a 42-nm diameter virion composed of an outer surface or envelope that carries the
hepatitis B surface antigen (HBsAg) (1, 10, 16). The envelope surrounds an inner core that contains the hepatitis B core
antigen (HBcAg) (3, 8, 14). Inside the core is the HBV-DNA genome. Another antigen, the hepatitis B e antigen (HBeAg),
is a viral core protein found in the bloodstream during active replication of HBV (19). HBsAg occurs in three different
modifications known as large HBsAg (LHBs), middle HBsAg (MHBs), and small HBsAg (SHBs), which is the main structural
component of the viral surface coat. In addition to these intact infectious particles, 22-nm diameter spherical and tubular
particles of various lengths are also observed in serum of persons infected with HBV in a proportion of 10 3-10 6 incomplete
particles to a mature virus. These defective particles consist only of an outer coat containing HBsAg, but no viral
nucleocapsid (HBcAg) and no nucleic acid (HBV-DNA).
HBsAg is a heterogeneous antigen. The principal determinant is called a and is common to all types of HBsAg: the region
contains eight cysteine residues which form disulphide bridges to maintain the correct conformation of the loop. Other major
determinants of the antigen are d/y and w/r. These determinant pairs are mutually exclusive, i.e., only the combinations adw,
adr, ayw, and ayr are possible (16, 17).
Most circulating antibodies are specific for the epitopes localized within the a loop amino acids. Mutations in this region
(amino acid substitution, insertion or deletion) can cause a conformational alteration which impairs the antibody-antigen
interaction, decreasing vaccine efficacy. In such cases infection occurs even in the presence of anti-HBs antibodies
(vaccine escape mutants). As a consequence, antibodies to the wild-type a determinant used in conventional HBsAg
diagnostic methods can fail to detect ongoing HBV infection (diagnostic escape mutants).
Hepatitis B diagnosis has been based on detection of serologic markers. Testing for these markers helps to determine the
presence of past or ongoing HBV infection, the acute or chronic stage of the disease, response to therapy, and/or the
immune status of the patient (4, 5, 9). HBsAg is the first serological marker to appear in the circulation, well before clinical
symptoms, and is the viral component usually found in the highest concentration in the serum of HBV-infected patients
(2, 5). The use of HBsAg and HBeAg quantitative assays has recently suggested that these serological markers may be
helpful in identifying patients likely to respond to anti-HBV treatments. Serum HBsAg levels are correlated with HBV
covalently-closed circular (ccc)DNA as well as with intrahepatic HBV-DNA. The significance of HBsAg in serum is
determined by evaluating it in relationship to the presence or absence of the other HBV markers and the clinical presentation
and history of the patient. HBsAg test, however, is of particular relevance in the screening of blood donations, for reducing
the incidence of post-transfusion HBV hepatitis.

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3. PRINCIPLE OF THE PROCEDURE
The method for quantitative determination of HBsAg is a direct two-step sandwich chemiluminescence immunoassay (CLIA).
Comparable sensitivity for detection of different mutants and genotypes is assured by mouse monoclonal antibodies directed
to highly conserved epitopes of HBsAg inner region that can detect HBsAg when used in combination with a complex
detergent mixture. Detergents used in the assay buffer make this approach manifest.
A mixture of mouse monoclonal antibodies is used for coating magnetic particles (solid phase) and a different mixture of
mouse monoclonal antibodies directed to different epitopes is linked to an isoluminol derivative (isoluminol-antibody
conjugate). During the first incubation, HBsAg present in calibrators, samples or controls binds to the solid phase.
During the second incubation, the antibody conjugate reacts with HBsAg already bound to the solid phase. After each
incubation, the unbound material is removed with a wash cycle.
Subsequently, the starter reagents are added and a flash chemiluminescence reaction is thus induced. The light signal, and
hence the amount of isoluminol-antibody conjugate, is measured by a photomultiplier as relative light units (RLU) and is
directly proportional to HBsAg concentration present in calibrators, samples or controls.

4. MATERIALS PROVIDED

Reagent integral
Magnetic particles [SORB] Magnetic particles coated with antibodies to HBsAg (mouse monoclonal) having balanced
(2.5 mL) reactivity for ad and ay subtypes, biotinylated BSA, streptavidin, BSA, PBS buffer, < 0.1%
sodium azide.
Calibrator 1 [CAL|1] Low levels of recombinant HBsAg (obtained in Hansenula polymorpha) having balanced
(3.0 mL) reactivity for ad and ay subtypes, BSA, phosphate buffer, EDTA, detergents, 0.2%
ProClin ® 300, an inert yellow dye. The calibrator concentrations (IU/mL) are referenced to
NIBSC standard (code 00/588, WHO Second International Standard for HBsAg, subtype
adw2, genotype A).
Calibrator 2 [CAL|2] High levels of recombinant HBsAg (obtained in Hansenula polymorpha) having balanced
(3.0 mL) reactivity for ad and ay subtypes, BSA, phosphate buffer, EDTA, detergents, 0.2%
ProClin ® 300, an inert blue dye. The calibrator concentrations (IU/mL) are referenced to
NIBSC standard (code 00/588, WHO Second International Standard for HBsAg, subtype
adw2, genotype A).
Buffer J [BUF|J] Non-specific IgG (mouse polyclonal), casein, urea, TRIS buffer, EDTA, detergents, 0.1%
(28 mL) ProClin ® 300.
Conjugate [CONJ] Mouse monoclonal IgG to HBsAg having balanced reactivity for ad and ay subtypes,
(2 x 21 mL) conjugated to an isoluminol derivative, human serum/plasma non-reactive for all HBV
markers, sheep serum, bovine serum, non-specific IgG (mouse polyclonal), BSA, phosphate
buffer, detergents, 0.2% ProClin ® 300, preservatives.
Number of tests 200
All reagents are supplied ready to use. The order of reagents reflects the layout of containers in the reagent integral.

Materials required but not provided

LIAISON ® XL Cuvettes ([REF] X0016).
LIAISON ® XL Disposable Tips ([REF] X0015).
LIAISON ® XL Starter Kit ([REF] 319200).
LIAISON ® Wash/System Liquid ([REF] 319100).
LIAISON ® XL Waste Bags ([REF] X0025).

Additionally required materials

LIAISON ® XL MUREX HBsAg Quant controls (negative and positive) ([REF] 310251).
LIAISON ® XL MUREX HBsAg Quant Specimen Diluent ([REF] 310252).

5. WARNINGS AND PRECAUTIONS

For in vitro diagnostic use.
All serum and plasma units used to produce the components provided in this kit have been tested for the presence of
HBsAg, anti-HCV, anti-HIV-1, anti-HIV-2 and found to be non-reactive, except for the positive control, which is reactive for
HBsAg. The hepatitis B surface antigen has been heat treated (60°C for 10 hours) during the manufacturing process.
Nevertheless, complete inactivation should not be assumed.
As, however, no test method can offer absolute assurance that pathogens are absent, all specimens of human origin should
be considered potentially infectious and handled with care.

6. SAFETY PRECAUTIONS
Do not eat, drink, smoke or apply cosmetics in the assay laboratory.
Do not pipette by mouth.
Avoid direct contact with potentially infected material by wearing laboratory clothing, protective goggles, and disposable
gloves. Wash hands thoroughly at the end of each assay.
Avoid splashing or forming an aerosol. All drops of biological reagent must be removed with a sodium hypochlorite solution
with 0.5% active chlorine, and the means used must be treated as infected waste.
All samples and reagents containing biological materials used for the assay must be considered as potentially able to
transmit infectious agents. The waste must be handled with care and disposed of in compliance with the laboratory
guidelines and the statutory provisions in force in each Country. Any materials for reuse must be appropriately sterilized in
compliance with the local laws and guidelines. Check the effectiveness of the sterilization/decontamination cycle.
Do not use kits or components beyond the expiration date given on the label.

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Pursuant to EC Regulation 1272/2008 (CLP) hazardous reagents are classified and labeled as follow:

REAGENTS: [CAL|1], [CAL|2], [CONJ], [BUF|J]

CLASSIFICATION: Acute Tox. 3 H331
Eye Dam. 1 H318
Skin sens. 1 H317
Skin Irrit. 2
Skin sens. 1 H317
SIGNAL WORD: Warning Danger
SYMBOLS /
PICTOGRAMS:

HAZARD STATEMENTS:
PRECAUTIONARY
STATEMENTS:
CONTAINS:
(only substances prescribed
pursuant to Article 18 of EC
Regulation 1272/2008).
GHS07 Exclamation mark
H317 May cause an allergic skin reaction.
P261 Avoid breathing dust/fume/gas/mist/
vapours/spray.
P280 Wear protective gloves/protective
clothing/eye protection/face protection.
P363 Wash contaminated clothing before
reuse.
reaction mass of: 5-chloro-2-methyl-
4-isothiazolin-3-one [EC no. 247-500-7]
and 2-methyl-2H -isothiazol-3-one
[EC no. 220-239-6] (3:1) (ProClin ® 300).
GHS05 Corrosion, GHS06 Skull and cross bones
H331 Toxic if inhaled;
H318 Causes serious eye damage;
H315 Causes skin irritation;
H317 May cause an allergic skin reaction.
P261 Avoid breathing dust/fume/gas/mist/
vapours/spray.
P280 Wear protective gloves/protective
clothing/eye protection/face protection.
P304+340 IF INHALED: remove victim to
fresh air and keep at rest in a position
comfortable for breathing;
P310 Immediately call a POISON CENTER
or doctor/physician
N-Lauroylsarcosine sodium salt
mixture of 5-chloro-2-methyl-2H-isothiazol-
3-one [EC No. 247-500-7];
2-methyl-2H-isothiazol-3-one
[EC No. 220-239-6] (3:1) (ProClin ® 300);
t-Octylphenoxypolyethoxyethanol.

Pursuant to EC Regulation 1272/2008 (CLP), [SORB] is labeled as EUH210 safety data sheets available on request.
For additional information see Safety Data Sheets available on www.diasorin.com.

7. REAGENT PREPARATION
REAGENT INTEGRAL
Please note the following important reagent handling precautions:
Resuspension of magnetic particles
Magnetic particles must be completely resuspended before the integral is placed on the instrument. Follow the
steps below to ensure complete suspension:
Before the seal is removed, rotate the small wheel at the magnetic particle compartment until the colour of the
suspension has changed to brown. Gentle and careful side-to-side mixing may assist in the suspension of the
magnetic particles (avoid foam formation). Visually check the bottom of the magnetic particle vial to confirm that
all settled magnetic particles have resuspended. Carefully wipe the surface of each septum to remove residual
liquid.
Repeat as necessary until the magnetic particles are completely resuspended.
An incomplete magnetic particles resuspension may cause variable and inaccurate analytical results.

Foaming of reagents
In order to ensure optimal performance of the integral, foaming of reagents should be avoided. Adhere to the recommendation
below to prevent this occurrence:
Visually inspect the reagents, calibrators in particular (position two and three following the magnetic particle vial), to ensure
there is no foaming present before using the integral. If foam is present after resuspension of the magnetic particles, place
the integral on the instrument and allow the foam to dissipate. Load the integral into the reagent area once the foam has
dissipated.
Loading of integral into the reagent area
– LIAISON ® XL Analyzer is equipped with a built-in solid-state magnetic device which aids in the dispersal of microparticles
prior to placement of a reagent integral into the reagent area of the analyzer. Refer to the analyzer operator's manual for
details.
a. Insert the reagent integral into the dedicated slot.
b. Allow the reagent integral to remain in the solid-state magnetic device for at least 30 seconds (up to several minutes).
Repeat as necessary.
– Place the integral into the reagent area of the analyzer with the label facing left and let it stand for 15 minutes before
using. The analyzer automatically stirs and completely resuspends the magnetic particles.
– Follow the analyzer operator's manual to load the specimens and start the run.
CONTROLS
Refer to the LIAISON ® XL MUREX HBsAg Quant Control Set instructions for use section for proper preparation and handling
instructions.

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8. REAGENT INTEGRAL STORAGE AND STABILITY
– Sealed: Stable at 2-8°C until the expiry date.
– Opened on board or at 2-8°C: Minimum stability four weeks.
After this period, it is still possible to keep on using the reagent integral provided that the controls are found within the
expected ranges.
– Use storage rack provided with the LIAISON ® XL Analyzer for upright storage of reagent integral.
– Do not freeze.
– Keep upright for storage to facilitate later proper resuspension of magnetic particles.
– Keep away from direct light.

9. SPECIMEN COLLECTION AND PREPARATION

Either human serum or plasma may be used (including serum collected in serum separator tubes). The anticoagulants sodium
citrate, potassium EDTA, lithium and sodium heparin, potassium oxalate, ACD (acid citrate-dextrose), CPDA
(citrate-phosphate-dextrose-adenine) have been tested and may be used with this assay. The correct specimen type must
be used in the assay.
Follow tube manufacturers’ instructions carefully when using collection containers. Blood should be collected aseptically by
venipuncture and the serum or plasma separated from clot, red cells or gel separator, after centrifugation.
Centrifugation conditions range from 1,000 to 3,000 g for 10 minutes. Conditions may vary depending on tube manufacturers
recommendations. Use of alternate centrifugation conditions should be evaluated and validated by the laboratory.
Before shipping specimens, serum or plasma specimens should be removed from clot, red cells or gel separator. Specimens
may be shipped in dry ice (frozen), in wet ice (for 2°-8°C) or at room temperature (20°-25°C), by following sample storage
limitations described below.
Uncontrolled transport conditions (in terms of temperature and time) can cause inaccurate analytical results. During
validation studies, specimen collection tubes commercially available at the time of testing were used. Therefore not all
collection tubes from all manufacturers have been evaluated. Blood collection devices from various manufacturers may
contain substances which could affect the test results in some cases (Bowen et al., Clinical Biochemistry, 43, 4-25, 2010).
Concerning storage limitations, if the assay is performed within seven days of sample collection, the samples removed from
red cells, clot or gel separator may be kept at 2°-8°C; otherwise they should be aliquoted and stored deep-frozen (–20°C or
below). Sixteen serum or plasma samples with different reactivity were stored for seven days at 2-8°C and 15 underwent four
freeze-thaw cycles. The results showed no significant differences; however multiple freeze-thaw cycles should be avoided. If
samples are stored frozen, mix thawed samples well before testing.
Samples removed from red cells, clot or gel separator having particulate matter, fibrin, turbidity, lipaemia, or erythrocyte
debris, specimens that have been stored at room temperature (20°-25°C), or frozen and thawed, or samples requiring repeat
testing, require clarification by further centrifugation (it’s recommended 10,000 g for 10’) before testing, to improve
consistency of results. Specimens with a lipid layer on the top should be transferred in a secondary tube, taking care to
transfer only the clarified material. Grossly haemolyzed or lipaemic samples as well as samples containing particulate matter
or exhibiting obvious microbial contamination should not be tested. Check for and remove air bubbles before assaying.
Cadaveric specimens should be stored following same indications than for living donors.
The minimum volume required for a single determination is 300 L specimen (150 L specimen + 150 L dead volume).

10. CALIBRATION
Test of assay specific calibrators allows the detected relative light unit (RLU) values to adjust the assigned master curve.
Each calibration solution allows five calibrations to be performed.
Recalibration in triplicate is mandatory whenever at least one of the following conditions occurs:
– A new lot of Starter Kit is used.
– The previous calibration was performed more than four weeks before.
– Each time a new lot of integral is used.
– The analyzer has been serviced.
– Control values lie outside the expected ranges.

11. ASSAY PROCEDURE

Strict adherence to the analyzer operator’s manual ensures proper assay performance. Each test parameter is identified via
information encoded in the reagent integral Radio Frequency IDentification transponder (RFID Tag). In the event that the
RFID Tag cannot be read by the analyzer, the integral cannot be used. Do not discard the reagent integral; contact your
local DiaSorin technical support for instruction.
The analyzer operations are as follows:
1. Dispense calibrators, controls or specimens into the reaction cuvettes.
2. Dispense buffer J.
3. Dispense coated magnetic particles.
4. Incubate.
5. Wash with Wash/System liquid.
6. Dispense conjugate into the reaction cuvettes.
7. Dispense buffer J.
8. Incubate.
9. Wash with Wash/System liquid.
10. Add the Starter Reagents and measure the light emitted.
Due to the presence of detergents in the LIAISON ® XL MUREX HBsAg Quant reagents, foam may be generated in the Liquid
Waste container. If this happens, in order to avoid overflow of the foam from the container it is advisable to empty the waste
container when the level of the liquid is approximately half of the capability of the container or alternatively to employ a
silicone based antifoam, to be added into the Liquid Waste container when it is empty and hypochlorite is added.

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12. QUALITY CONTROL
LIAISON ® XL controls should be run in singlicate to monitor the assay performance. Quality control must be performed by
running LIAISON ® XL MUREX HBsAg Quant controls
(a) at least once per day of use,
(b) whenever a new reagent integral is used,
(c) whenever the kit is calibrated,
(d) whenever a new lot of Starter Reagents is used,
(e) to assess adequacy of performance of the open integral beyond four weeks, or in agreement with guidelines or
requirements of local regulations or accredited organizations.
Control values must lie within the expected ranges: whenever one or both controls lie outside the expected ranges,
calibration should be repeated and controls retested. If control values obtained after successful calibration lie repeatedly
outside the predefined ranges, the test should be repeated using an unopened control vial. If control values lie outside the
expected ranges, patient results must not be reported.
The performance of other controls should be evaluated for compatibility with this assay before they are used. Appropriate value
ranges should then be established for quality control materials used.

13. INTERPRETATION OF RESULTS

The analyzer automatically calculates HBsAg concentrations expressed as IU/mL and grades the results. For details, refer
to the analyzer operator’s manual.
Assay range. 0.030 to 150 IU/mL HBsAg.
Specimens containing HBsAg concentrations above the assay range (above 150 IU/mL) may be automatically diluted using
LIAISON® XL MUREX HBsAg Quant Specimen Diluent ([REF] 310252) loaded in the ancillary reagent area.
The recommended dilution factor is 1:400; dilution should never exceed 1:999. The dilution factor should be chosen in order
that the result after dilution scores above 0.05 IU/mL.
The cut-off value discriminating between the presence and the absence of HBsAg is 0.05 IU/mL. Sample results should be
interpreted as follows:
Samples with HBsAg concentration values below 0.05 IU/mL should be graded non-reactive.
Samples with HBsAg concentration values equal to or above 0.05 IU/mL should be graded reactive.
A sample resulting reactive at the first assay should be assayed again in duplicate. If a sample results repeatedly reactive in
at least one replicate, the characterization of individual’s HBV infection should be evaluated with supplementary investigation,
such as other HBV markers or HBV DNA or HBsAg Confirmatory Test, [REF]310110. Samples which are non-reactive at the
second test should be considered non-reactive.

14. LIMITATIONS OF THE PROCEDURE

A skillful technique and strict adherence to the instructions are necessary to obtain reliable results.
Bacterial contamination or heat inactivation of the specimens may affect the test results.
Specimens should be kept at room temperature only for the amount of time required for handling and preparation.
This test is suitable only for investigating single samples, not sample pools.
Falsely reactive results may be obtained with any diagnostic test. Two kinds of falsely reactive result may be observed with
LIAISON ® XL MUREX HBsAg Quant test: non-reproducibly reactive results and non-specifically reactive results.
Non-reproducibly reactive results may be caused by contamination of the reaction cuvettes caused by trace amounts of
extremely high-positive samples processed immediately before a negative specimen. This potential interference, however,
does not affect the integrity of the original test tube. As a consequence, falsely reactive samples are eventually correctly
classified, when reactive specimens are retested in duplicate in accordance with the retesting algorithm recommended by
the assay protocol.
Specimens from individuals recently vaccinated against HBV may score transiently positive for HBsAg because it is present
in the vaccine. Reactivity to vaccine may vary with different manufacturers' tests.
Non-specifically reactive results may be observed in most highly sensitive immunoassays. For instance, specimens from
patients receiving preparations of mouse monoclonal antibodies for therapy or diagnosis may contain human anti-mouse
antibodies (HAMA). Such specimens may interfere in a monoclonal antibody-based immunoassay. Non-specifically reactive
results, however, are eventually correctly classified by HBsAg Confirmatory Test, [REF] 310110.
Test results are reported quantitatively as positive or negative for the presence of HBsAg. However, an HBsAg-negative
result does not exclude the possibility of exposure to or infection with hepatitis B virus. Diagnosis of infectious diseases
should not be established on the basis of a single test result, but should be determined in conjunction with clinical findings
and other diagnostic procedures as well as in association with medical judgement. A full differential diagnostic work-up for
the diagnosis of hepatitis B and related clinical conditions includes examination of the patient's immune status and clinical
history.
Specimens from patients receiving therapeutic doses of Biotin (Vitamin H, B7 or B8) may interfere in immunoassays based
on biotinylated reagents. Interference was not observed testing Biotin serum concentration up to 3500 ng/mL with
LIAISON ® XL Murex HBsAg Quant assay (for details, refer to §15.1).
Before testing cadaveric specimens, collection and centrifugation procedures should be carefully applied. After death,
haemolysis and other changes (including proteolysis and dilution) occur in blood, which may lead to False Negative and
False Positive in testing. In subjects transfused immediately prior to death high percentage of haemodilution can affect the
performance of the test due to analyte dilution.

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15. SPECIFIC PERFORMANCE CHARACTERISTICS

15.1. Analytical specificity

Analytical specificity may be defined as the ability of the assay to accurately detect specific analyte in the presence of
potentially interfering factors in the sample matrix (e.g., anticoagulants, haemolysis, effects of sample treatment), or
cross-reactive antibodies.
Interference. Controlled studies of potentially interfering substances or conditions showed that the assay performance was
not affected by anticoagulants (sodium citrate, potassium EDTA, lithium and sodium heparin, potassium oxalate, ACD, CPDA),
haemolysis (up to 1000 mg/dL haemoglobin), lipaemia (up to 3000 mg/dL triglycerides), bilirubinaemia (up to 20 mg/dL
bilirubin), serum levels of Biotin (up to 3500 ng/mL) or by a limited number of freeze-thaw cycles of samples. Results are not
influenced by the use of positive same-day fresh samples as a comparative study in 30 freshly collected specimens
demonstrates.
Cross-reactions. The cross-reactivity study for the LIAISON ® XL MUREX HBsAg Quant assay was designed to evaluate
potential interference from antibodies to other organisms that may cause infectious diseases (EBV, hCMV, rubella virus,
parvovirus B19, Toxoplasma gondii, Treponema pallidum, Borrelia burgdorferi, HSV, VZV, HAV, HIV, HCV) as well as from
other conditions that may result from atypical immune system activity (anti-nuclear autoantibodies, rheumatoid factor, human
anti-mouse antibodies). Samples for these studies were pre-screened with another commercially available HBsAg assay.
If found negative for HBsAg, those specimens were used to study potential cross-reactivity. The presence of potential
cross-reactants in the samples was detected using CE-marked assays.
Number of expected LIAISON ® XL
Condition negative samples positive results
hCMV IgG antibodies 10 0
EBV (VCA) IgG antibodies 15 0
HSV-1/2 IgG antibodies 11 0
Rubella virus IgG antibodies 15 0
Parvovirus B19 IgG antibodies 15 0
VZV IgG antibodies 14 0
HCV antibodies 6 0
HIV antibodies and p24 antigen 14 0
HAV antibodies 7 0
HTLV-I/II antibodies 8 0
Borrelia burgdorferi IgG antibodies 7 0
Toxoplasma gondii IgG antibodies 10 0
Treponema pallidum antibodies 16 0
Rheumatoid factor (anti-Fc immunoglobulin) 8 0
Anti-nuclear autoantibodies (ANA) 10 0
Human anti-mouse antibodies (HAMA) 20 0
Total 186 0

15.2. Analytical sensitivity

Analytical sensitivity may be expressed as the limit of detection (LoD), which is the minimal amount of specific analyte
detectable by the assay. The limit of detection was calculated in accordance with the guidelines of Clinical and Laboratory
Standards Institute (CLSI, USA), document No. EP17-A.
60 negative samples were tested in singlicate with three kit lots on two instruments and the results were used to calculate
the limit of blank (LoB) as the 95th percentile on each lot. Then five samples containing low HBsAg concentrations were
tested in 20 runs with the same three kit lots and two instruments. The obtained results allowed calculation of the limit of
detection of each kit, as follows: LoD = LoB + 1.645 x S.D., where S.D. is the pooled standard deviation.
The LIAISON ® XL MUREX HBsAg Quant assay analytical sensitivity (LoD) is less than or equal to 0.030 IU/mL.
The study performed with the Second International Standard for HBsAg, subtype adw2, genotype A, NIBSC code: 00/588,
showed a sensitivity of 0.05 IU/mL.
15.3. Precision
Different samples, containing different concentrations of specific analyte along the assay range, were assayed to estimate
repeatability and reproducibility of the assay (i.e., within- and between-assay variability). The results refer to the groups of
samples investigated and are not guaranteed specifications, as differences may exist between laboratories and locations.
Repeatability. Twenty replicates were performed in the same run to evaluate in-house repeatability.
Positive
Repeatability A B C D E
control
Number of determinations 20 20 20 20 20 20
Mean (IU/mL) 0.33 0.51 0.53 11.15 82.00 0.24
Standard deviation (IU/mL) 0.02 0.04 0.05 0.49 9.82 0.01
Coefficient of variation (%) 6.09 7.29 9.69 4.39 11.98 6.34
Min. value (IU/mL) 0.28 0.40 0.34 10 64 0.22
Max. value (IU/mL) 0.37 0.55 0.57 12 94 0.27

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Reproducibility. Twenty determinations were performed in different days (one or two runs per day) with three different lots
of integral to evaluate reproducibility. The tests were performed using two instruments.
Positive
Reproducibility - Instrument 1 A B C D E
control
LOT No. 01
Number of determinations 20 20 20 20 20 20
Mean (IU/mL) 0.32 0.54 0.57 12.16 84.83 0.28
Standard deviation (IU/mL) 0.03 0.03 0.04 0.76 8.64 0.02
Coefficient of variation (%) 9.07 5.09 6.73 6.21 10.19 6.52
Min. value (IU/mL) 0.27 0.48 0.48 11 61 0.26
Max. value (IU/mL) 0.38 0.56 0.63 14 95 0.31
LOT No. 02
Number of determinations 20 20 20 20 20 20
Mean (IU/mL) 0.25 0.43 0.47 11.36 80.74 0.21
Standard deviation (IU/mL) 0.02 0.02 0.03 0.66 9.15 0.02
Coefficient of variation (%) 8.19 5.68 6.82 5.84 11.33 8.60
Min. value (IU/mL) 0.23 0.39 0.37 11 57 0.18
Max. value (IU/mL) 0.31 0.47 0.51 13 89 0.26
LOT No. 03
Number of determinations 20 20 20 20 20 20
Mean (IU/mL) 0.28 0.48 0.50 11.56 80.69 0.23
Standard deviation (IU/mL) 0.02 0.02 0.03 0.53 10.11 0.01
Coefficient of variation (%) 5.97 3.23 6.35 4.55 12.54 4.70
Min. value (IU/mL) 0.25 0.44 0.41 11 46 0.21
Max. value (IU/mL) 0.31 0.51 0.54 12 92 0.25
Inter-lot coefficient of variation (%) 13.02 10.42 10.40 6.23 11.41 13.62

Positive
Reproducibility - Instrument 2 A B C D E
control
LOT No. 01
Number of determinations 20 20 20 20 20 20
Mean (IU/mL) 0.29 0.51 0.53 11.43 77.24 0.25
Standard deviation (IU/mL) 0.03 0.02 0.04 0.98 8.11 0.01
Coefficient of variation (%) 9.80 4.77 6.99 8.64 10.50 5.98
Min. value (IU/mL) 0.23 0.46 0.43 9.5 56 0.23
Max. value (IU/mL) 0.34 0.55 0.60 13 89 0.28
LOT No. 02
Number of determinations 20 20 20 20 20 20
Mean (IU/mL) 0.27 0.48 0.51 11.75 84.66 0.23
Standard deviation (IU/mL) 0.03 0.03 0.03 0.93 7.12 0.03
Coefficient of variation (%) 9.86 6.06 6.24 7.90 8.41 14.08
Min. value (IU/mL) 0.23 0.41 0.46 9.4 68 0.20
Max. value (IU/mL) 0.34 0.54 0.59 13 97 0.36
LOT No. 03
Number of determinations 20 20 20 20 20 20
Mean (IU/mL) 0.26 0.45 0.48 11.36 80.18 0.22
Standard deviation (IU/mL) 0.02 0.02 0.02 0.85 7.54 0.01
Coefficient of variation (%) 7.23 4.85 5.07 7.49 9.40 4.12
Min. value (IU/mL) 0.23 0.40 0.44 10 63 0.20
Max. value (IU/mL) 0.30 0.49 0.52 13 91 0.23
Inter-lot coefficient of variation (%) 10.19 6.81 7.60 8.02 10.01 10.81

15.4. Trueness
The assay trueness has been checked by the dilution test.
Dilution test. Four serum samples containing high HBsAg concentrations were tested as such and after serially diluting with
the specimen diluent. HBsAg concentrations measured versus concentrations expected were analyzed by linear regression.
The correlation coefficients (r) ranged from 0.959 to 1.000.
Expected Measured Expected Measured
Dilution concentration, concentration, % Recovery Dilution concentration, concentration, % Recovery
IU/mL IU/mL IU/mL IU/mL
neat – 26,400 – neat – 6,800 –
1:200 132.0 120 90.9 1:100 68.0 58 85.3
1:400 66.0 63 95.5 1:200 34.0 35 102.9
1:800 33.0 31 93.9 1:400 17.0 17 100.0
1:1600 16.5 20 121.0 1:800 8.5 8.6 101.2
neat – 14,200 – neat – 13,600 –
1:100 142.0 110 77.5 1:100 136.0 130 95.6
1:200 71.0 75 105.6 1:200 68.0 64 94.1
1:400 35.5 34 95.8 1:400 34.0 32 94.1
1:800 17.8 18 101.4 1:800 17.0 16 94.1

15.5. High-dose saturation effect

Whenever samples containing extremely high antigen concentrations are tested, the saturation effect can mimic
concentrations lower than real. However, a well-optimized two-step method excludes grossly underestimated results,
because the analytical signals remain consistently high (saturation curve).
Analysis of saturation effect was evaluated by testing seven high-titred samples positive for HBsAg. All samples resulted in
estimated concentration values above the assay range that would be expected with high-titred samples, indicating no
sample misclassification.

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15.6. Performance characteristics of cadaveric specimen testing
Performance characteristics of cadaveric specimens testing was determined by testing, according PEI validation protocol*,
post-mortem specimens collected up to 24 hours after death in comparison to living donor specimens. 40 post-mortem
samples were tested as unspiked and spiked at 2 levels: low positive and medium/high positive. The same procedure was
performed with the same number of normal human serum from living donors, tested in parallel as reference to compare with
post-mortem sample results. The results obtained were analyzed through calculation of percentage difference between
mean of living donors results and mean of post-mortem results, at each reactivity level. In this study, the obtained
percentage difference was equal or below 4,6% for each of the tested reactivity levels (see table below). Paired t-test
analysis were performed between post-mortem and living donors specimens, spiked at low and medium/high positive levels,
demonstrating not significantly difference on two groups (p value <0.05).
Repeteability was assessed using one post-mortem and one living donor specimens, spiked up to a low-level of reactivity
with a human serum reactive for antibodies to hepatitis C virus (HCV). Each specimen was assessed in six replicates in the
same run. The obtained percent coefficient of variation (CV%) did not exceed 15%. As reported in the table below 5.3% for the
cadaveric specimen and 5.1% for the living donor were found in the study. T he results refer to t he group of investigated sa
mples and are not guaranteed specifications, as differences may exist between laboratories and locations.
Test results Recovery (%) t-test CV%
Sample
Means (IU/mL) Post-mortem/Living donors p value 6 replicates
Post-Mortem unspiked 0.03
Neat n.a. n.a n.a
Living donors unspiked 0.03

Post-Mortem spiked 0.15 5.3

Low Positive -4.6 0.129
Living donors spiked 0.15 5.1

Post-Mortem spiked 0.30

Medium/high Positive 3.3 0.297 n.a
Living donors spiked 0.29

* Paul Ehrlich Institute - Proposal for the Validation of Anti-HIV-1/2 or HIV Ag/Ab Combination Assays, Anti-HCV-Assays,
HBsAg and Anti-HBc Assays for Use with Cadaveric Samples - 08/05/2014

16. EXPECTED VALUES

Diagnostic specificity and sensitivity were estimated in accordance with the updated version of Common Technical
Specification (CTS) published on Nov. 27, 2009 (Art. 5, §3 of IVD Directive 98/79/EC). The results refer to the groups of
samples investigated and are not guaranteed specifications, as differences may exist between laboratories and locations.
16.1. Diagnostic specificity
A study was performed on a total of 5,201 serum and plasma specimens collected in two blood donation centres (including
100 specimens from first-time donors). Specimens tested were expected negative samples from an unselected blood donor
population with zero prevalence of HBV infection. The assay shows diagnostic specificity above 99.5% (95% confidence
interval: 99.89-100%). Additional specimens were also tested, randomly selected from hospitalized patients, dialysis
patients, pregnant women, high-risk subjects (i.e., haemophiliacs, intravenous drug users, homosexual males, and patients
affected by sexually-transmitted diseases). Data of these studies are summarized in Table I (95% CI = 95% confidence
interval). Positive specimens were confirmed by a reference CE-marked kit.

Table I - Diagnostic specificity.

Initially Repeat Confirmed Diagnostic
Number Diagnostic
Population reactive reactive positive specificity,
of cases specificity, %
samples, No. samples, No. samples, No. 95% CI
Blood donors 5201 6 1 0 99.98 (5200/5201) 99.89-100.0
Hospitalized patients 390 5 4 4 100.0 (386/386) 99.05-100.0
Dialysis patients 278 3 3 2 99.64 (275/276) 98.00-100.0
Pregnant women 100 0 0 0 100.0 (100/100) 96.38-100.0
High-risk subjects 143 7 5 4 99.28 (138/139) 96.05-99.98

16.2. Diagnostic sensitivity

Diagnostic sensitivity was assessed by testing 424 specimens from preselected HBsAg-positive patients (86 of whom with
defined HBsAg subtypes). Diagnostic sensitivity of this s tudy is 100% (95% confidence interval: 99.1-100.0%).
Besides, results obtained are in agreement with those expected in 22 samples with defined HBsAg subtypes selected from
DiaSorin repository panel as well as four commercially available panels encompassing different HBsAg subtypes (ad and ay)
and genotypes.
In an additional study the ability of the LIAISON ® XL MUREX HBsAg Quant assay to detect HBsAg was evaluated by testing
sequentially-collected specimens belonging to 30 seroconversion panels from donors who seroconverted over the course of
their donation history. Commercially available, precharacterized panels for HBV antigens were used, each starting with a
negative bleed and exhibiting narrow bleeding intervals. The panels were also tested by a reference CE-marked HBsAg
assay. The results show that the LIAISON ® XL MUREX HBsAg Quant assay detected HBsAg one bleed earlier in five out of
30 panels. The reference assay detected HBsAg one bleed earlier in two out of 30 panels. Both assays exhibited equivalent
HBsAg detection in 23 out of 30 panels.
The test diagnostic sensitivity in the detection of HBV early infection is therefore substantially equivalent to the reference
assay.

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16.3. Mutant HBsAg detection
Table II shows data about HBsAg mutant susceptibility obtained by testing ten common mutants, including the most prevalent
Gly–Arg 145 mutant (G145R). The panel was also tested by reference CE-marked HBsAg assays (A and B).

Table II - Diagnostic sensitivity in mutant HBsAg detection.

Liaison® XL Liaison ® XL
Mutant MUREX HBsAg Assay A Assay B Mutant MUREX HBsAg Assay A Assay B
Quant, IU/mL Quant, IU/mL
Mutant 1 0.92 (+) reactive reactive Mutant 6 1.60 (+) reactive non-reactive
Mutant 2 0.24 (+) non-reactive non-reactive Mutant 7 0.36 (+) reactive reactive
Mutant 3 0.31 (+) non-reactive reactive Mutant 8 0.53 (+) reactive reactive
Mutant 4 0.81 (+) reactive reactive Mutant 9 0.50 (+) reactive reactive
Mutant 5 1.00 (+) reactive reactive Mutant 10 0.45 (+) reactive reactive

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Changes: -
Deletions: -

LIAISON ® XL HBsAg Quant Specimen Diluent ([REF] 310252)

1. Intended use.
LIAISON ® XL MUREX HBsAg Quant Specimen Diluent ([REF] 310252) is to be used to dilute specimens containing HBsAg
concentrations above the assay range (above 150 IU/mL) of LIAISON ® XL MUREX HBsAg Quant assay. Refer to §15.4 of
instructions for use for detailed data on trueness by dilution test. The performance characteristics of the diluent have not
been established for any other assays or instrument platforms different from LIAISON ® XL.
2. Materials provided

Specimen diluent (50 mL) [DIL|SPE] Human serum/plasma non-reactive for all HBV markers, stabilized in TRIS buffer,
0.2% ProClin ® 300, preservatives.
The diluent is supplied ready to use. It is not kit lot specific and may be safely used even with different reagent integral lots.
In order to maintain reagent traceability on the instrument, do not separate Specimen Diluent bottle from the ancillary holder.
3. Safety precautions
Do not eat, drink, smoke or apply cosmetics in the assay laboratory.
Do not pipette by mouth.
Avoid direct contact with potentially infected material by wearing laboratory clothing, protective goggles, and disposable
gloves. Wash hands thoroughly at the end of each assay.
Avoid splashing or forming an aerosol. All drops of biological reagent must be removed with a sodium hypochlorite solution
with 0.5% active chlorine, and the means used must be treated as infected waste.
All samples and reagents containing biological materials used for the assay must be considered as potentially able to
transmit infectious agents. The waste must be handled with care and disposed of in compliance with the laboratory
guidelines and the statutory provisions in force in each Country. Any materials for reuse must be appropriately sterilized in
compliance with the local laws and guidelines. Check the effectiveness of the sterilization/decontamination cycle.
Do not use kits or components beyond the expiration date given on the label.
Pursuant to EC Regulation 1272/2008 (CLP) hazardous reagents are classified and labeled as follow:

REAGENTS: [DIL|SPE]
CLASSIFICATION Skin sens. 1 H317
SIGNAL WORD: Warning
SYMBOLS / PICTOGRAMS:

GHS07 Exclamation mark

HAZARD STATEMENTS: H317 May cause an allergic skin reaction.
PRECAUTIONARY STATEMENTS: P261 Avoid breathing dust/fume/gas/mist/vapours/spray.
P280 Wear protective gloves/protective clothing/eye protection/face protection.
P363 Wash contaminated clothing before reuse.

CONTAINS:
reaction mass of: 5-chloro-2-methyl-4-isothiazolin-3-one [EC no. 247-500-7] and
(only substances prescribed pursuant to 2-methyl-2H -isothiazol-3-one [EC no. 220-239-6] (3:1) (ProClin® 300).
Article 18 of EC Regulation 1272/2008).

For additional information see Safety Data Sheets available on www.diasorin.com.

4. Reagent preparation. At the time of use, equilibrate the specimen diluent to room temperature (20-25°C) then open the
vial. The specimen diluent must be loaded onto the instrument in the ancillary reagent area. After use, recap the vial
promptly and store at 2-8°C in an upright position.
Each specimen diluent vial is identified via information encoded in the reagent integral Radio Frequency IDentification
transponder (RFID Tag). In case the RFID Tag cannot be read, the specimen diluent vial cannot be used and must be
discarded.
For details on the reagent use in the ancillary reagent area on board the instrument, refer to the LIAISON ® XL operator’s
manual.
5. Storage and stability. Upon receipt, the specimen diluent must be stored at 2-8°C in an upright position to prevent
adherence of the solution to the vial cap. Do not freeze. When the diluent is stored sealed and kept upright, it is stable at
2-8°C up to the expiry date. Once opened the diluent is stable for four weeks when properly stored at 2-8°C between two
successive uses. Avoid bacterial contamination. The diluent should not be used past the expiry date indicated on the vial
label.

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Changes: §1;
Deletions: -

LIAISON ® XL Control HBsAg Quant ([REF] 310251)

1. INTENDED USE
The LIAISON ® XL MUREX HBsAg Quant controls (negative and positive) are intended for use as assayed quality control
samples to monitor the performance and reliability of the LIAISON ® XL MUREX HBsAg Quant assay. The performance
characteristics of LIAISON ® XL MUREX HBsAg Quant controls have not been established for any other assays or instrument
platforms different from LIAISON ® XL.
The certificate of analysis bar codes give specific information on the lot of controls and should be read by the hand-held bar
code scanner of the LIAISON ® XL Analyzer prior to loading the control vials on board. For details, refer to the analyzer
operator's manual.

2. MATERIALS PROVIDED

Negative control (2 x 4.0 mL) [CONTROL|-] Human serum/plasma non-reactive for all HBV markers, stabilized in TRIS
buffer, 0.2% ProClin ® 300, preservatives.

Positive control (2 x 4.0 mL) [CONTROL|+] Human serum/plasma reactive for hepatitis B surface antigen (ad and ay),
stabilized in TRIS buffer, 0.2% ProClin ® 300, preservatives.
All reagents are supplied ready to use. The range of concentrations of each control is reported on the certificate of analysis
and indicates the limits established by DiaSorin for control values that can be obtained in reliable assay runs. Each
laboratory is responsible for adopting different limits to meet individual requirements.

3.WARNINGS AND PRECAUTIONS

– For in vitro diagnostic use.
– Controls are not kit lot specific and may be safely interchanged even with different reagent integral lots.
– All materials used to produce the components provided in this kit have been tested for the presence of HBsAg, anti-HCV,
anti-HIV-1, anti-HIV-2 and found to be non-reactive, except for the positive control, which is reactive for HBsAg. The
hepatitis B surface antigen has been heat treated (60°C for 10 hours) during the manufacturing process. Nevertheless,
complete inactivation should not be assumed.
As, however, no test method can offer absolute assurance that pathogens are absent, all specimens of human origin
should be considered potentially infectious and handled with care.
– Observe the normal precautions required for handling all laboratory reagents.
– Disposal of all waste material should be in accordance with local guidelines.

4. SAFETY PRECAUTIONS
Do not eat, drink, smoke or apply cosmetics in the assay laboratory.
Do not pipette by mouth.
Avoid direct contact with potentially infected material by wearing laboratory clothing, protective goggles, and disposable
gloves. Wash hands thoroughly at the end of each assay.
Avoid splashing or forming an aerosol. All drops of biological reagent must be removed with a sodium hypochlorite solution
with 0.5% active chlorine, and the means used must be treated as infected waste.
All samples and reagents containing biological materials used for the assay must be considered as potentially able to
transmit infectious agents. The waste must be handled with care and disposed of in compliance with the laboratory
guidelines and the statutory provisions in force in each Country. Any materials for reuse must be appropriately sterilized in
compliance with the local laws and guidelines. Check the effectiveness of the sterilization/decontamination cycle.
Do not use kits or components beyond the expiration date given on the label.
Pursuant to EC Regulation 1272/2008 (CLP) hazardous reagents are classified and labeled as follow:

REAGENTS: [CONTROL|-], [CONTROL|+]

CLASSIFICATION: Skin sens. 1 H317
SIGNAL WORD: Warning
SYMBOLS / PICTOGRAMS:

GHS07 Exclamation mark

HAZARD STATEMENTS:
PRECAUTIONARY STATEMENTS:
CONTAINS:
(only substances prescribed pursuant to
Article 18 of EC Regulation 1272/2008).
H317 May cause an allergic skin reaction.
P261 Avoid breathing dust/fume/gas/mist/vapours/spray.
P280 Wear protective gloves/protective clothing/eye protection/face protection.
P363 Wash contaminated clothing before reuse.
reaction mass of: 5-chloro-2-methyl-4-isothiazolin-3-one [EC no. 247-500-7] and
2-methyl-2H -isothiazol-3-one [EC no. 220-239-6] (3:1). (ProClin® 300).

For additional information see Safety Data Sheets available on www.diasorin.com.

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5. STORAGE AND STABILITY
Upon receipt, the controls must be stored at 2-8°C in an upright position to prevent adherence of the solution to the vial cap.
Do not freeze. When controls are stored sealed and kept upright, they are stable at 2-8°C up to the expiry date.
Once opened controls are stable for four weeks when properly stored at 2-8°C between two successive uses. Avoid bacterial
contamination of controls. The controls should not be used past the expiry date indicated on the vial labels.

6. PREPARATION OF REAGENTS
– Place the control vials in type C racks on the LIAISON ® XL Analyzer. Each control solution allows at least 20 tests to be
performed.
– The minimum volume required is 550 L (150 L control + 400 L dead volume).
– At the time of use, equilibrate controls to room temperature (20-25°C) before opening the vials and keep them on board
the instrument only for the amount of time required for quality control testing.
– After use, stopper the vials promptly and store them at 2-8°C in an upright position.
– During handling, use appropriate precautions to avoid bacterial contamination of controls.

7. HANDLING
For proper handling please refer to the LIAISON ® XL Analyzer operator’s manual.

8. TARGET VALUES
The target values and ranges of HBsAg concentrations in the controls are printed on the certificate of analysis. They have
been established after taking into account run variability with respect to the stored master curve, in order to guarantee
accuracy of analytical results and to obtain indications on stability or deterioration of reagents. If controls values lie repeatedly
outside the expected ranges, the test has most probably been performed incorrectly.

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REFERENCES

1. B.S. BLUMBERG, H.J. ALTER

A “new” antigen in leukemia sera.
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Electron microscopy of hepatitis B core antigen synthesized in E. coli.
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Hepatitis B.
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9. W.H. GERLICH, R. THOMSSEN

Terminology, structure, and laboratory diagnosis of hepatitis viruses.
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Large surface proteins of hepatitis B virus containing the pre-S sequence.
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Chronic viral hepatitis.
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Understanding hepatitis B virus infection.
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Comparison of the amino acid sequences of nine different serotypes of hepatitis surface antigen and genomic classification of the corresponding
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Relationship between serotypes and genotypes of hepatitis B virus: genetic classification of HBV by use of surface genes.
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18. M. RIZZETTO, G. LANFRANCO, R. PAGNI, G. VERME
The diagnostic approach to chronic hepatitis.
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p. 3-13 (1995).
19. H.J. SCLICHT, J. SALFELD, H. SCHALLER
The duck hepatitis B virus pre-C region encodes a signal sequence which is essential for synthesis and secretion of processed core proteins but
not for virus formation.
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20. W. SZMUNESS
Recent advances in the study of the epidemiology of hepatitis B.
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Genetic variations of the hepatitis B virus and their clinical relevance.
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Additional References
T.F. BAUMERT et al.
Pathogenesis of hepatitis B virus infection.
World J. Gastroenterol., 13 (1) : 82-90 (2007).
M.R. BRUNETTO, U.A. RODRIGUEZ, F. BONINO
Hepatitis B virus mutants.
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EUROPEAN PATENT Application EP 1 806 363 A1, 2007; Bulletin 2007/28.
S. IJAZ et al.
A 'first loop' linear epitope accessible on native hepatitis B surface antigen that persists in the face of 'second loop' immune escape.
J. Gen. Vir., 84 : 269-275 (2003).
J.M. JONGERIUS et al.
New hepatitis B virus mutant form in a blood donor that is undetectable in several hepatitis B surface antigen screening assays.
Transfusion, 38 : 56-59 (1998).
E. KAJIWARA et al.
Hepatitis B caused by a hepatitis B surface antigen escape mutant.
J. Gastroenterol., 43 : 243-247 (2008).
S. LEVICNIK-STEZINAR
Hepatitis B surface antigen escape mutant in a first time blood donor potentially missed by a routine screening assay.
Clin. Lab., 50 : 49-51 (2004).
T.D. LY et al.
Sensitivities of four new commercial hepatitis B virus surface antigen (HBsAg) assays in detection of HBsAg mutant forms.
J. Clin. Microbiol., 44 (7) : 2321-2326 (2006).
T.D. LY
Detection of HBsAg mutants by immunoassays.
J. Med. Virol., 79 : S37-S41 (2007).
K.M. WEINBERGER et al.
High genetic variability of the group-specific a-determinant of hepatitis B virus surface antigen (HBsAg) and the corresponding fragment of the viral
polymerase in chronic virus carriers lacking detectable HBsAg in serum.
J. Gen. Virol., 81 : 1165-1174 (2000).
A. Krawczyk, C. Hintze, J. Ackermann, B. Goitowski, M. Trippler, N. Grüner, M. Neumann-Fraune, J. Verheyen and M. Fiedler
Clinical performance of the novel DiaSorin LIAISON ® XL murex: HBsAg Quant, HCV-Ab, HIV-Ab/Ag assays.
J Clin Virol.59 : 44-49 (2014).

E. Burdino, T. Ruggiero, A. Proietti, M.G. Milia, A. Olivero, G.P. Caviglia, M. Marietti, M. Rizzetto, A. Smedile, V. Ghisetti
Quantification of hepatitis B surface antigen with the novel DiaSorin LIAISON ® XL Murex HBsAg Quant: Correlation with the ARCHITECT quantitative
assays.
J Clin Virol.60 : 341-346 (2014).

K. Malm, E. Kragsbjerg and S. Andersson

Performance of Liaison XL automated immunoassay platform for blood-borne infection screening on hepatitis B, hepatitis C, HIV 1/2, HTLV 1/2 and
Treponema pallidum serological markers.
Transfusion Medicine 25 (2) : 101-105 (2015).

P. Grimse, N. Frey, G. Bending, J. Zitzler, O. Lorenz, D. Kasapic & C. E Zaugg

Population pharmacokinetics of exogenous biotin and the relationship between biotin serum levels and in vitro immonoassay interference.
International Journal of Pharmacokinetics
(2017) 2(4); 247-256

Additional References for use of cadaveric samples

Proposal for the Validation of Anti-HIV-1/2 or HIV Ag/Ab Combination Assays, Anti-HCV-Assays, HBsAg and Anti-HBc Assays for Use with Cadaveric
Samples - 08/05/2014.
C. BALERIOLA et al.
Infectious disease screening of blood specimens collected post-mortem provides comparable results to pre-mortem specimens.
Cell Tissue Bank (2012) 13; page 251-258.
WE FINKBEINER, P URSELL, RL DAVIS
Autopsy Pathology: A Manual and Atlas (2004), Cap 9; page 113-118.
FL DELMONICO
Cadaver donor screening for infectious agents in solid organ transplantation.
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200/007-926, 07- 2019-10

HBsAgQ-rf.fm LIAISON® XL MUREX HBsAg Quant ([REF] 310250)

200/007-926, 07- 2019-10
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