Elecsys Anti-HBc II

REF SYSTEM
MODULAR ANALYTICS E170
cobas e 411
07394764 119 200
cobas e 601
cobas e 602

English Reagents – working solutions

System information The reagent rackpack (M, R0, R1, R2) is labeled as A-HBCII.
For cobas e 411 analyzer: test number 1460 M Streptavidin-coated microparticles (transparent cap), 1 bottle, 12 mL:
For MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers: Streptavidin-coated microparticles 0.72 mg/mL; preservative.
Application Code Number 715
Intended Use R0 DTT (white cap), 1 bottle, 5 mL:
Immunoassay for the in vitro qualitative determination of IgG and IgM 1,4-dithiothreitol 110 mmol/L; citrate buffer 50 mmol/L.
antibodies to the hepatitis B core antigen in human serum and plasma. R1 HBcAg (gray cap), 1 bottle, 15 mL:
This assay is indicated as an aid in the diagnosis of infection with HBV. This HBcAg (E. coli, rDNA) > 25 ng/mL; phosphate buffer 100 mmol/L,
assay is also indicated as a donor screening test to detect HBV infection in
serum or plasma specimens from individual human blood donors. It may also pH 7.4; preservative.
be used in testing serum or plasma specimens to screen individual organ R2 Anti-HBcAg-Ab~biotin; anti-HBcAg-Ab~Ru(bpy)2+ 3 (black cap),
donors when specimens are obtained while the donor's heart is still beating.
1 bottle, 15 mL:
The electrochemiluminescence immunoassay “ECLIA” is intended for use on
Elecsys and cobas e immunoassay analyzers. Biotinylated monoclonal anti-HBc antibody (mouse) 700 ng/mL;
Note: Please note that the catalogue number appearing on the package monoclonal anti-HBc antibody (mouse) labeled with ruthenium complex
insert retains only the first 8 digits of the licensed 11-digit Catalogue Number: 200 ng/mL; phosphate buffer 100 mmol/L, pH 7.4; preservative.
07394764190 for the Elecsys Anti-HBc II assay. The last 3 digits -190 have
been replaced by -119 for logistic purposes. A-HBCII Cal1 Negative calibrator 1 (white cap), 2 bottles of 1.0 mL:
Summary each:
The hepatitis B virus (HBV) consists of an external envelope (HBsAg) and an Human serum; preservative.
inner core (HBcAg). The hepatitis core antigen comprises 183-185 amino
acids.1 A-HBCII Cal2 Positive calibrator 2 (black cap), 2 bottles of 1.0 mL each:
During an infection with HBV, antibodies to HBcAg are generally produced, Anti-HBc (human) > 8 WHO IU/mLb) in human serum;
which often persist for life. Anti-HBc appears shortly after the onset of HBV preservative.
infection and can usually be detected in serum soon after the appearance of
HBsAg.2 Anti-HBc persists in persons who have recovered from HBV b) WHO international units
infection and inactive carriers. Therefore, they are an indicator of existing or
past HBV infection.1,2,3 Precautions and warnings
In rare cases, HBV infection can also run its course without the appearance For in vitro diagnostic use.
of immunologically detectable anti-HBc (usually in immunosuppressed Exercise the normal precautions required for handling all laboratory reagents.
patients). Anti-HBc is not produced after vaccination.4 Disposal of all waste material should be in accordance with local guidelines.
Safety data sheet available for professional user on request.
Due to the persistence of anti-HBc following infection with HBV, screening for
anti-HBc can be used to identify previously infected individuals.5 All human material should be considered potentially infectious. All products
derived from human blood are prepared exclusively from the blood of donors
Determination of anti-HBc in association with other hepatitis B tests permits tested individually and shown to be free from HBsAg (A-HBCII Cal1 only) and
the diagnosis and monitoring of HBV infections. In the absence of other antibodies to HCV and HIV. The testing methods used assays approved by
hepatitis B markers (HBsAg-negative persons), anti-HBc may be the only the FDA or cleared in compliance with the European Directive 98/79/EC,
indication of an existing HBV infection.2,6,7 Annex II, List A.
Test principle The serum containing anti-HBc (A-HBCII Cal2) was inactivated using
Competition principle. Total duration of assay: 27 minutes. ß-propiolactone and UV-radiation.
 1st incubation: Pretreatment of 40 µL of sample with reducing agent. However, as no inactivation or testing method can rule out the potential risk
of infection with absolute certainty, the material should be handled with the
 2nd incubation: After addition of HBcAg, a complex is formed with anti- same level of care as a patient specimen. In the event of exposure, the
HBc antibodies in the sample. directives of the responsible health authorities should be followed.8,9
 3rd incubation: After addition of biotinylated antibodies and ruthenium Avoid foam formation in all reagents and sample types (specimens,
complexa) -labeled antibodies specific for HBcAg, together with calibrators and controls).
streptavidin-coated microparticles, the still-free binding sites on the HBc- Reagent Handling
antigens become occupied. The entire complex becomes bound to the
solid phase via interaction of biotin and streptavidin. The reagents in the kit are ready-for-use and are supplied in bottles
compatible with the system.
 The reaction mixture is aspirated into the measuring cell where the cobas e 411 analyzer: The calibrators should only be left on the analyzer
microparticles are magnetically captured onto the surface of the during calibration at 20-25 °C. After use, close the bottles as soon as
electrode. Unbound substances are then removed with ProCell/ possible and store upright at 2-8 °C.
ProCell M. Application of a voltage to the electrode then induces
chemiluminescent emission which is measured by a photomultiplier. Due to possible evaporation effects, not more than 5 calibration procedures
per bottle set should be performed.
 Results are determined automatically by the software by comparing the MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers:
electrochemiluminescence signal obtained from the reaction product of Unless the entire volume is necessary for calibration on the analyzers,
the sample with the signal of the cutoff value previously obtained by transfer aliquots of the ready-for-use calibrators into empty snap-cap bottles
calibration. (CalSet Vials). Attach the supplied labels to these additional bottles. Store the
a) Tris(2,2’-bipyridyl)ruthenium(II)-complex (Ru(bpy)2+
3 )
aliquots at 2-8 °C for later use.
Perform only one calibration procedure per aliquot.

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Elecsys Anti-HBc II
All information required for correct operation is read in from the respective ▪ General laboratory equipment
reagent barcodes. ▪ MODULAR ANALYTICS E170 or cobas e analyzer
Please note: Both the vial labels, and the additional labels (if available) Accessories for cobas e 411 analyzer:
contain 2 different barcodes. The barcode between the yellow markers is
for cobas 8000 systems only. If using a cobas 8000 system, please turn ▪ 11662988122, ProCell, 6 x 380 mL system buffer
the vial cap 180° into the correct position so the barcode can be read by the ▪ 11662970122, CleanCell, 6 x 380 mL measuring cell cleaning
system. Place the vial on the instrument as usual. solution
Storage and stability ▪ 11930346122, Elecsys SysWash, 1 x 500 mL washwater additive
Store at 2‑8 °C. ▪ 11933159001, Adapter for SysClean
Do not freeze.
▪ 11706802001, AssayCup, 60 x 60 reaction cups
Store the Elecsys reagent kit upright in order to ensure complete
availability of the microparticles during automatic mixing prior to use. ▪ 11706799001, AssayTip, 30 x 120 pipette tips
▪ 11800507001, Clean‑Liner
Stability of the reagent rackpack Accessories for MODULAR ANALYTICS E170, cobas e 601 and
unopened at 2‑8 °C up to the stated expiration date cobas e 602 analyzers:
after opening at 2‑8 °C 8 weeks ▪ 04880340190, ProCell M, 2 x 2 L system buffer
on the analyzers 8 weeks ▪ 04880293190, CleanCell M, 2 x 2 L measuring cell cleaning
solution
Stability of the calibrators ▪ 03023141001, PC/CC‑Cups, 12 cups to prewarm ProCell M and
CleanCell M before use
unopened at 2‑8 °C up to the stated expiration date
▪ 03005712190, ProbeWash M, 12 x 70 mL cleaning solution for run
after opening at 2‑8 °C 8 weeks finalization and rinsing during reagent change
on cobas e 411 at 20‑25 °C up to 6 hours ▪ 03004899190, PreClean M, 5 x 600 mL detection cleaning solution
on MODULAR ANALYTICS E170, use only once ▪ 12102137001, AssayTip/AssayCup, 48 magazines x 84 reaction
cobas e 601 and cobas e 602 at cups or pipette tips, waste bags
20‑25 °C ▪ 03023150001, WasteLiner, waste bags
Store calibrators upright in order to prevent the calibrator solution from ▪ 03027651001, SysClean Adapter M
adhering to the snap‑cap. Accessories for all analyzers:
Specimen collection and preparation ▪ 11298500316, ISE Cleaning Solution/Elecsys SysClean,
Only the specimens listed below were tested and found acceptable. 5 x 100 mL system cleaning solution
Serum collected using standard sampling tubes or tubes containing Assay
separating gel. For optimum performance of the assay follow the directions given in this
Li‑heparin, Na‑heparin, K2‑EDTA, K3‑EDTA, ACD, CPD, CP2D, CPDA and document for the analyzer concerned. Refer to the appropriate operator’s
Na‑citrate plasma. manual for analyzer‑specific assay instructions.
Plasma tubes containing separating gel can be used. Resuspension of the microparticles takes place automatically prior to use.
Criterion: Correct assignment of positive and negative samples. Samples Read in the test‑specific parameters via the reagent barcode. If in
with a COI (cutoff index) > 1.0: ± 20 % recovery; samples with a COI ≤ 1.0: exceptional cases the barcode cannot be read, enter the 15‑digit sequence
± 0.20 recovery. of numbers (except for the cobas e 602 analyzer).
Stable for 7 days at 15‑25 °C, 14 days at 2‑8 °C, 3 months at ‑20 °C MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers:
(± 5 °C). The samples may be frozen 5 times. PreClean M solution is necessary.
The sample types listed were tested with a selection of sample collection Bring the cooled reagents to approximately 20 °C and place on the reagent
tubes or systems that were commercially available at the time of testing, i.e. disk (20 °C) of the analyzer. Avoid foam formation. The system
not all available tubes of all manufacturers were tested. Sample collection automatically regulates the temperature of the reagents and the
systems from various manufacturers may contain differing materials which opening/closing of the bottles.
could affect the test results in some cases. When processing samples in Place the calibrators in the sample zone.
primary tubes (sample collection systems), follow the instructions of the All the information necessary for calibrating the assay is automatically read
tube manufacturer. into the analyzer.
Attention: Particularly important for the Elecsys Anti‑HBc II assay: Thawed After calibration has been performed, store the calibrators at 2‑8 °C or
samples, samples containing precipitates, and samples for repeat discard (MODULAR ANALYTICS E170, cobas e 601 and cobas e 602
measurements must be carefully centrifuged before performing the assay. analyzers).
The performance of the Elecsys Anti‑HBc II assay has not been established
with cadaveric samples or body fluids other than serum and plasma. Calibration
Traceability: This method has been standardized against the anti-HBc
Do not use samples and controls stabilized with azide. reference material WHO Standard (NIBSC code 95/522).
Ensure the samples, calibrators and controls are at 20‑25 °C prior to Calibration frequency: Calibration must be performed once per reagent lot
measurement. using A‑HBCII Cal1, A‑HBCII Cal2 and fresh reagent (i.e. not more than
Due to possible evaporation effects, samples and calibrators on the 24 hours since the reagent kit was registered on the analyzer).
analyzers should be analyzed/measured within 2 hours. Calibration interval may be extended based on acceptable verification of
Materials provided calibration by the laboratory.
See “Reagents – working solutions” section for reagents. Renewed calibration is recommended as follows:
▪ 2 x 4 bottle labels ▪ after 1 month (28 days) when using the same reagent lot
Materials required (but not provided) ▪ after 7 days (when using the same reagent kit on the analyzer)
▪ 04927931190, PreciControl Anti‑HBc II, for 8 x 1.3 mL ▪ as required: e.g. quality control findings with PreciControl Anti‑HBc II
▪ 11776576322, CalSet Vials, 2 x 56 empty snap‑cap bottles outside the defined limits

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Elecsys Anti-HBc II
Range for the electrochemiluminescence signals (counts) for the Limits and ranges
calibrators: Detection limit: ≤ 0.8 WHO IU/mL
Negative calibrator (A‑HBCII Cal1):
42000‑300000 (cobas e 411 analyzer) The stated sensitivity was determined by reading off the anti‑HBc
100000‑700000 (MODULAR ANALYTICS E170, cobas e 601 and concentration corresponding to the signal of the cutoff value from standard
cobas e 602 analyzers) curves obtained by serial dilution of the WHO anti‑HBc reference material in
Positive calibrator (A‑HBCII Cal2): human serum free from hepatitis B.
100‑3000 (MODULAR ANALYTICS E170 and cobas e analyzers) Specific performance data
Quality control Representative performance data on the analyzers are given below.
For quality control, use PreciControl Anti‑HBc II. Results obtained in individual laboratories may differ.
Controls for the various concentration ranges should be run individually at Precision
least once every 24 hours when the test is in use, once per reagent kit, and Precision was determined using Elecsys reagents, human sera and
following each calibration. controls.
The control intervals and limits should be adapted to each laboratory’s
individual requirements. Values obtained should fall within the defined cobas e 411 analyzer
limits. Each laboratory should establish corrective measures to be taken if Repeatabilityc) Intermediate
values fall outside the defined limits. precisiond)
Follow the applicable government regulations and local guidelines for
quality control. Sample Mean SD CV SD CV
COI COI % COI %
If necessary, repeat the measurement of the samples concerned.
Note: HSe), negative 2.23 0.029 1.3 0.070 3.2
For technical reasons re-assigned target values valid only for a specific HS, weakly positive 0.855 0.016 1.9 0.034 3.9
reagent and control lot combination must be entered manually on all
analyzers (except for the cobas e 602 analyzer). Therefore always refer to HS, positive 0.029 0.001 4.0 0.002 6.6
the value sheet included in the reagent kit or PreciControl kit to make sure PCf) Anti‑HBc II 1 1.92 0.039 2.0 0.094 4.9
that the correct target values are used.
When a new reagent or control lot is used, the analyzer will use the original PC Anti‑HBc II 2 0.490 0.012 2.5 0.039 8.0
values encoded in the control barcodes. c) Precision according to CLSI EP05-A3 (21 days) (Clinical and Laboratory Standards Institute)
Calculation d) Intermediate precision according to CLSI EP05-A3 (21 days) (Clinical and Laboratory
The analyzer automatically calculates the cutoff based on the measurement Standards Institute)
of A‑HBCII Cal1 and A‑HBCII Cal2. e) HS = human serum
The result of a sample is given either as reactive or non-reactive as well as f) PC = PreciControl
in the form of a cutoff index (signal sample/cutoff). MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers
Interpretation of the results Repeatability Intermediate
Samples with a cutoff index > 1.0 are non‑reactive in the Elecsys precision
Anti‑HBc II assay. These samples are considered negative for anti‑HBc and
do not need further testing. Sample Mean SD CV SD CV
Samples with a cutoff index ≤ 1.0 are reactive in the Elecsys Anti‑HBc II COI COI % COI %
assay. All samples found to be reactive in the initial test must be retested in HS, negative 2.30 0.042 1.8 0.062 2.7
duplicate with the Elecsys Anti‑HBc II assay. If the results in the follow-up
test are “non-reactive” in both cases, then the sample is deemed negative HS, weakly positive 0.882 0.019 2.2 0.043 4.9
for anti‑HBc. HS, positive 0.028 0.0008 2.8 0.002 7.3
If at least one of the repeat measurements is reactive, then the sample is
deemed repeatedly reactive. PC Anti‑HBc II 1 2.22 0.034 1.5 0.065 2.9
Limitations - interference PC Anti‑HBc II 2 0.603 0.019 3.2 0.032 5.4
The assay is unaffected by icterus (bilirubin ≤ 1129 µmol/L or ≤ 66 mg/dL), Analytical specificity
hemolysis (Hb ≤ 0.621 mmol/L or ≤ 1000 mg/dL), lipemia (Intralipid
≤ 2000 mg/dL) and biotin (≤ 123 nmol/L or ≤ 30 ng/mL). 309 samples containing potentially interfering substances were tested with
the Elecsys Anti‑HBc II assay comprising specimens:
Criterion:
▪ containing antibodies against HAV, HCV, HIV, HSV, Rubella, CMV,
Samples > 1.0 COI: ± 20 % recovery EBV, Toxoplasma gondii, Treponema pallidum
Samples ≤  1.0 COI: ± 0.20 recovery
▪ positive for E. coli
Samples should not be taken from patients receiving therapy with high
biotin doses (i.e. > 5 mg/day) until at least 8 hours following the last biotin ▪ after vaccination against HAV and HBV
administration. ▪ non-viral induced liver diseases
No interference was observed from rheumatoid factors up to a ▪ autoimmune diseases (ANA and SLE)
concentration of 1200 IU/mL. No false reactive results were found with the Elecsys Anti‑HBc II assay
No significant interfering effects of 24 commonly used and special (after repetition) resulting in a specificity of 100 %. 81 samples were
therapeutic drugs could be detected. Special drugs tested: pegylated identified as congruently positive with the Elecsys Anti‑HBc II and another
interferon alpha‑2 a, pegylated interferon alpha‑2 b, lamivudine, adefovir, commercially available Anti‑HBc assay. 1 sample was found to be
entecavir, telbivudin, tenofovir. indeterminate and was excluded from calculation.
In rare cases, interference due to extremely high titers of antibodies to Clinical sensitivity
analyte‑specific antibodies, streptavidin or ruthenium can occur. These
effects are minimized by suitable test design. Of 793 samples from HBV infected patients in different stages of the
disease, 793 were found repeatedly reactive with the Elecsys Anti‑HBc II
For diagnostic purposes, the results should always be assessed in assay. The sensitivity of the Elecsys Anti‑HBc II assay in this study was
conjunction with the patient’s medical history, clinical examination and other found to be 100 %.
findings.

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Elecsys Anti-HBc II
Stage of disease N Reactive 7 Guidelines for vaccinating kidney dialysis patients and patients with
chronic kidney disease. Summarized from recommendations of the
Chronic or acute HBV infection (anti-HBc positive, 568 568 advisory committee on immunization practices (ACIP). December 2012.
HBsAg positive) http://www.cdc.
Passed HBV infection (anti-HBc positive, HBsAg 56 56 gov/dialysis/PDFs/Vaccinating_Dialysis_Patients_and_Patients_dec20­
12.pdf
negative, anti-HBs positive, anti-HBe negative)
8 Occupational Safety and Health Standards: Bloodborne pathogens.
Passed or recovered HBV infection (anti-HBc 9 9 (29 CFR Part 1910.1030). Fed. Register.
positive, HBsAg negative, anti-HBs negative, anti-
9 Directive 2000/54/EC of the European Parliament and Council of
HBe positive) 18 September 2000 on the protection of workers from risks related to
Passed or recovered HBV infection (anti-HBc 160 160 exposure to biological agents at work.
positive, HBsAg negative, anti-HBs positive, anti- For further information, please refer to the appropriate operator’s manual for
HBe positive) the analyzer concerned, the respective application sheets, the product
information and the Method Sheets of all necessary components (if
Seroconversion sensitivity available in your country).
Seroconversion sensitivity of the Elecsys Anti‑HBc II assay has been A point (period/stop) is always used in this Method Sheet as the decimal
investigated by testing 10 commercially available seroconversion panels. separator to mark the border between the integral and the fractional parts of
Elecsys Anti‑HBc II assay was shown to be sensitive in early detection of a decimal numeral. Separators for thousands are not used.
infection in line with other anti‑HBc assays and additional HBV serological
markers. Symbols
Roche Diagnostics uses the following symbols and signs in addition to
Clinical specificity those listed in the ISO 15223‑1 standard (for USA: see
A total of 20101 samples from blood donors, diagnostic routine, pregnant https://usdiagnostics.roche.com for definition of symbols used):
women and dialysis patients from 7 centers in Europe was tested with the
Elecsys Anti‑HBc II assay. Contents of kit
The resulting specificity in the study based on a repeatedly reactive basis Analyzers/Instruments on which reagents can be used
(RR) was 99.93 % in blood donors (serum), the 95 % confidence interval
(2‑sided) was 99.84‑99.97 %. Reagent
Specificity in blood donors (plasma) was found to be 99.88 %, the 95 % Calibrator
confidence interval (2‑sided) was 99.78‑99.94 %.
Volume after reconstitution or mixing
Specificity in diagnostic routine/hospitalized patients was found to be 100 %
(95 % confidence interval (2‑sided): 99.60‑100 %), in dialysis patients GTIN Global Trade Item Number
99.31 % (95 % confidence interval (2‑sided): 98.23‑99.81 %) and in
pregnant women 100 % (95 % confidence interval (2‑sided): 99.62‑100 %). COBAS, COBAS E, ELECSYS and PRECICONTROL are trademarks of Roche. INTRALIPID is a trademark of
Fresenius Kabi AB.
Cohort N Specificity (%) 95 % confidence Specificity (%) 95 % confidence
All other product names and trademarks are the property of their respective owners.
IRg) interval (2‑sided) RR interval (2‑sided) Additions, deletions or changes are indicated by a change bar in the margin.
(%) IR (%) RR © 2017, Roche Diagnostics
Blood donors 8163 99.93 99.84-99.97 99.93 99.84-99.97
(serum)

Blood donors 9162 99.88 99.78-99.94 99.88 99.78-99.94

Roche Diagnostics GmbH, Sandhofer Strasse 116, D-68305 Mannheim
(EDTA plasma) www.roche.com

Diagnostic 997 100 99.60-100 100 99.60-100

routine / hospital­
ized patients

Dialysis patients 779 99.13 97.99-99.72 99.31 98.23-99.81

Pregnant women 1000 100 99.62-100 100 99.62-100

g) IR = initially reactive
References
1 Seeger C, Zoulim F, Mason WS. Hepadnaviruses. In: Field’s Virology,
Knipe DM, Howley RM (eds), 2007 5th edition, Lippincott Williams and
Wilkins, Philadelphia, USA. Chapter 76, pp2977-3029.
2 Liaw YF, Chu CM. Hepatitis B virus infection. Lancet
2009;373:582-592.
3 Elgouhari HM, Abu-Rajab Tamini TI, Carey W. Hepatitis B virus
infection: understanding its epidemiology, course, and diagnosis. Cleve
Clin J Med 2008;75:881-889.
4 Caspari G, Gerlick WH. The serologic markers of hepatitis B virus
infection – proper selection and standardized interpretation. Clin Lab
2007;53:335-343.
5 WHO. Hepatitis B. WHO/CDS/CSR/LYO/2002.2:Hepatitis B. Available
at: http://www.who.int/csr/disease/hepatitis/
HepatitisB_whocdscsrlyo2002_2.pdf, accessed February 2012.
6 Pondé RA, Cardoso DD, Ferro MO. The underlying mechanisms for the
‘anti-HBc alone’ serological profile. Arch Virol 2010;155:149-158.

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