07026838500V4.

Elecsys Anti-HBe
cobas e 402
07026838190 07026838500 300
cobas e 801

English ▪ The reaction mixture is aspirated into the measuring cell where the
microparticles are magnetically captured onto the surface of the
System information electrode. Unbound substances are then removed with ProCell II M.
Short name ACN (application code number) Application of a voltage to the electrode then induces chemiluminescent
emission which is measured by a photomultiplier.
AHBE 10033
▪ Results are determined automatically by the software by comparing the
Intended use electrochemiluminescence signal obtained from the reaction product of
Immunoassay for the in vitro qualitative determination of human antibodies the sample with the signal of the cutoff value previously obtained by
to the hepatitis B e antigen (HBeAg) in human serum and plasma. calibration.
a) Tris(2,2'-bipyridyl)ruthenium(II)-complex (Ru(bpy) )
The electrochemiluminescence immunoassay “ECLIA” is intended for use
on cobas e immunoassay analyzers. Reagents - working solutions
Regulatory approval The cobas e pack (M, R1, R2) is labeled as AHBE.
This assay has been CE marked according to Directive 98/79/EC. Test M Streptavidin-coated microparticles, 1 bottle, 12.4 mL:
performance has been established and certified by a Notified Body Streptavidin-coated microparticles 0.72 mg/mL; preservative.
according to the Common Technical Specifications (CTS) for diagnostic use
and for testing of blood donations. R1 HBeAg, 1 bottle, 21.9 mL:
Summary HBeAg (E. coli, rDNA) > 7 ng/mL; HEPESb) buffer 36 mmol/L, pH 7.4;
Hepatitis B virus (HBV) is transmitted by percutaneous or mucosal preservative.
exposure to infected blood and various body fluids including saliva, R2 Anti-HBeAg-Ab~biotin; anti-HBeAg-Ab~Ru(bpy) , 1 bottle, 13.9 mL:
menstrual, vaginal, and seminal fluids.1 The majority of adult patients Biotinylated monoclonal anti‑HBe antibody (mouse) > 0.8 mg/L;
recover completely from their HBV infection, but up to 10 % of the cases
become asymptomatic carriers or develop chronic hepatitis which may lead monoclonal anti‑HBe antibody (mouse) labeled with ruthenium
to cirrhosis and/or liver cancer.2,3 Despite immunization, HBV is still complex > 0.2 mg/L; HEPES buffer 36 mmol/L, pH 7.4; preservative.
prevalent worldwide with approximately 250 million chronically infected b) HEPES = [4-(2-hydroxyethyl)-piperazine]-ethane sulfonic acid
patients and a serious threat to blood transfusion safety, especially in highly
endemic countries.4,5 AHBE Cal1 Negative calibrator 1, 1 bottle of 1.0 mL:
Serological diagnosis of HBV infection involves the detection of HBV Human serum, preservative.
specific antigens and/or antibodies to identify different phases of the HBV
infection to determine whether a patient has acute or chronic HBV infection, AHBE Cal2 Positive calibrator 2, 1 bottle of 1.0 mL:
is susceptible to infection, or is immune to HBV as a result of prior infection Anti‑HBe (human) approximately 3 IU/mL in human serum;
or vaccination.6,7 Some of these HBV markers are also routinely used in preservative.
patient and donor screening.7
Precautions and warnings
The hepatitis B e antigen (HBeAg) is a product of the pre‑C/C gene that has
been found in hepatocytes during proliferation of the hepatitis B virus (HBV) For in vitro diagnostic use.
and is an important diagnostic tool to determine the status of ongoing HBV Exercise the normal precautions required for handling all laboratory
infections. The detection of HBeAg is generally associated with the reagents.
presence of large quantities of virus as it is a surrogate of viral replication.8,9 Disposal of all waste material should be in accordance with local guidelines.
During acute HBV infection HBeAg can be detected in serum shortly after Safety data sheet available for professional user on request.
hepatitis B surface antigen (HBsAg) and usually disappears before HBsAg, This kit contains components classified as follows in accordance with the
when alanine aminotransferase (ALT) levels peak, followed by the presence Regulation (EC) No. 1272/2008:
of the corresponding antibody (anti‑HBe).8,9,10 HBeAg can usually be
detected when viral replication is high; its presence for more than 10 weeks
is indicative of a persistent infection. HBeAg seroconversion to anti‑HBe
suggests the end of active viral replication and is therefore associated with
clinical resolution (self-limited) or remission (chronic disease).6,8,9,11 HBV
infections can occur without detectable HBeAg due to infection with HBV
variants containing precore stop codon mutants; while the virus can no Warning
longer produce HBeAg, disease activity is ongoing and anti‑HBe may be H317 May cause an allergic skin reaction.
present.8,12,13
The anti‑HBe test, therefore, is meaningful in association with the HBeAg Prevention:
test for monitoring the course of a HBV infection and the effect of treatment
for chronic hepatitis B.6,8,9,11 The Elecsys Anti‑HBe assay uses recombinant P261 Avoid breathing dust/fume/gas/mist/vapours/spray.
HBeAg and monoclonal anti‑HBe antibodies to detect anti‑HBe.
P272 Contaminated work clothing should not be allowed out of
Test principle the workplace.
Competition principle. Total duration of assay: 18 minutes.
▪ 1st incubation: Anti‑HBe in the sample (21 µL) binds to the added P280 Wear protective gloves.
HBeAg. Response:
▪ 2nd incubation: After addition of biotinylated antibodies and ruthenium
complexa)-labeled antibodies specific for HBeAg, together with P333 + P313 If skin irritation or rash occurs: Get medical
streptavidin-coated microparticles, the still-free binding sites on the advice/attention.
HBe‑antigens become occupied. The entire complex is then bound to
the solid phase via interaction of biotin and streptavidin. P362 + P364 Take off contaminated clothing and wash it before reuse.
Disposal:

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Elecsys Anti-HBe
P501 Dispose of contents/container to an approved waste primary tubes (sample collection systems), follow the instructions of the
disposal plant. tube/collection system manufacturer.
Product safety labeling follows EU GHS guidance. Centrifuge samples containing precipitates, thawed samples, and samples
for repeat measurements before performing the assay. Heat-inactivated
Contact phone: all countries: +49-621-7590 samples may be used.
All human material should be considered potentially infectious. Do not use samples and controls stabilized with azide.
The negative calibrator (AHBE Cal1) has been prepared exclusively from Ensure the samples and calibrators are at 20‑25 °C prior to measurement.
the blood of donors tested individually and shown to be free from HBsAg
and antibodies to HCV and HIV. Due to possible evaporation effects, samples and calibrators on the
analyzers should be analyzed/measured within 2 hours.
The testing methods used assays approved by the FDA or cleared in
compliance with the European Directive 98/79/EC, Annex II, List A. The performance of the Elecsys Anti‑HBe assay has not been established
with cadaveric samples or body fluids other than serum and plasma.
The positive calibrator (AHBE Cal2) containing anti‑HBe was tested for HIV
and hepatitis C infections. The findings were negative. The serum Materials provided
containing anti‑HBe was inactivated using β‑propiolactone and See “Reagents – working solutions” section for reagents.
UV‑radiation.
▪ 2 x 4 bottle labels
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 Materials required (but not provided)
same level of care as a patient specimen. In the event of exposure, the ▪ 11876384122, PreciControl Anti‑HBe, for 16 x 1.3 mL
directives of the responsible health authorities should be followed.14,15
▪ 11776576322, CalSet Vials, 2 x 56 empty snap-cap bottles
Avoid foam formation in all reagents and sample types (specimens,
calibrators and controls). ▪ General laboratory equipment
Reagent handling ▪ cobas e analyzer
The reagents (M, R1, R2) in the kit are ready-for-use and are supplied in Additional materials for cobas e 402 and cobas e 801 analyzers:
cobas e packs. ▪ 06908799190, ProCell II M, 2 x 2 L system solution
Calibrators ▪ 04880293190, CleanCell M, 2 x 2 L measuring cell cleaning
The calibrators are supplied ready‑for‑use in bottles compatible with the solution
system. ▪ 07485409001, Reservoir Cup, 8 cups to supply ProCell II M and
Unless the entire volume is necessary for calibration on the analyzer, CleanCell M
transfer aliquots of the ready‑for‑use calibrators into empty snap-cap bottles ▪ 06908853190, PreClean II M, 2 x 2 L wash solution
(CalSet Vials). Attach the supplied labels to these additional bottles. Store
the aliquots at 2‑8 °C for later use. ▪ 05694302001, Assay Tip/Assay Cup tray, 6 magazines
Perform only one calibration procedure per aliquot. x 6 magazine stacks x 105 assay tips and 105 assay cups, 3 wasteliners
All information required for correct operation is available via the cobas link. ▪ 07485425001, Liquid Flow Cleaning Cup, 2 adaptor cups to supply
ISE Cleaning Solution/Elecsys SysClean for Liquid Flow Cleaning
Storage and stability Detection Unit
Store at 2‑8 °C. ▪ 07485433001, PreWash Liquid Flow Cleaning Cup, 1 adaptor cup
Do not freeze. to supply ISE Cleaning Solution/Elecsys SysClean for Liquid Flow
Store the cobas e pack upright in order to ensure complete availability of Cleaning PreWash Unit
the microparticles during automatic mixing prior to use. ▪ 11298500316, ISE Cleaning Solution/Elecsys SysClean,
5 x 100 mL system cleaning solution
Stability of the cobas e pack:
Assay
unopened at 2‑8 °C up to the stated expiration date For optimum performance of the assay follow the directions given in this
on the analyzers 16 weeks document for the analyzer concerned. Refer to the appropriate operator’s
manual for analyzer‑specific assay instructions.
Stability of the calibrators: Resuspension of the microparticles takes place automatically prior to use.
unopened at 2‑8 °C up to the stated expiration date Place the cooled (stored at 2‑8 °C) cobas e pack on the reagent manager.
Avoid foam formation. The system automatically regulates the temperature
after opening at 2‑8 °C 16 weeks of the reagents and the opening/closing of the cobas e pack.
on the analyzers at 20‑25 °C use only once Calibrators:
Store calibrators upright in order to prevent the calibrator solution from Place the calibrators in the sample zone.
adhering to the snap‑cap. Read in all the information necessary for calibrating the assay.
Specimen collection and preparation Calibration
Only the specimens listed below were tested and found acceptable. Traceability: This method has been standardized against the WHO 1st
Serum collected using standard sampling tubes or tubes containing International Standard Anti‑hepatitis B virus e antigen (anti‑HBe),
separating gel. code 129095/12 of the Paul‑Ehrlich‑Institut, Langen (Germany).
Li‑heparin, Na‑heparin, K2‑EDTA, K3‑EDTA, ACD, CPD, CP2D, CPDA and Calibration frequency: Calibration must be performed once per reagent lot
Na‑citrate plasma. using AHBE Cal1, AHBE Cal2 and fresh reagent (i.e. not more than
24 hours since the cobas e pack was registered on the analyzer).
Criterion: Samples with a COI (cutoff index) > 1.0: ± 20 % recovery;
samples with a COI ≤ 1.0: ± 0.20 recovery. Renewed calibration is recommended as follows:
Stable for 7 days at 20‑25 °C, 14 days at 2‑8 °C, 3 months at ▪ after 12 weeks when using the same reagent lot
‑20 °C (± 5 °C). The samples may be frozen 6 times. ▪ after 28 days when using the same cobas e pack on the analyzer
The sample types listed were tested with a selection of sample collection ▪ as required: e.g. quality control findings with PreciControl Anti‑HBe
tubes or systems that were commercially available at the time of testing, i.e. outside the defined limits
not all available tubes of all manufacturers were tested. Sample collection
systems from various manufacturers may contain differing materials which Range for the electrochemiluminescence signals (counts) for the
could affect the test results in some cases. When processing samples in calibrators:

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Elecsys Anti-HBe
Negative calibrator (AHBE Cal1): 300000‑1500000 Drug Concentration tested
Positive calibrator (AHBE Cal2): 1000‑6000 mg/L
Quality control Entecavir ≤1
For quality control, use PreciControl Anti‑HBe.
Telbivudine ≤ 600
Controls for the various concentration ranges should be run individually at
least once every 24 hours when the test is in use, once per cobas e pack, Tenofovir ≤ 245
and following each calibration. In rare cases, interference due to extremely high titers of antibodies to
The control intervals and limits should be adapted to each laboratory’s analyte‑specific antibodies, streptavidin or ruthenium can occur. These
individual requirements. Values obtained should fall within the defined effects are minimized by suitable test design.
limits. Each laboratory should establish corrective measures to be taken if For diagnostic purposes, the results should always be assessed in
values fall outside the defined limits. conjunction with the patient’s medical history, clinical examination and other
If necessary, repeat the measurement of the samples concerned. findings.
Follow the applicable government regulations and local guidelines for Limits and ranges
quality control.
Detection limit: < 0.2 IU/mL
Calculation The stated sensitivity was determined by reading off the anti‑HBe
The analyzer automatically calculates the cutoff based on the measurement concentration corresponding to the signal of the cutoff value from standard
of AHBE Cal1 and AHBE Cal2. The result of a sample is given either as curves obtained by serial dilution of the WHO anti‑HBe reference material in
reactive or non-reactive as well as in the form of a cutoff index (signal human serum free from hepatitis B.
sample/cutoff).
Expected values
Interpretation of the results Anti‑HBe could be detected in samples from 210 (83.7 %) out of
Numeric result Result message Interpretation/ 251 persons with chronic or past HBV infections. 14 (1.4 %) out of
1000 samples of randomly selected blood donors were reactive for
further steps anti‑HBe.
COI > 1.0 Non-reactive Negative for anti‑HBe, no Each laboratory should investigate the transferability of the expected values
further testing needed. to its own patient population and if necessary determine its own reference
ranges.
COI ≤ 1.0 Reactive Positive for anti‑HBe.
Specific performance data
Limitations - interference Representative performance data on the analyzers are given below.
The effect of the following endogenous substances and pharmaceutical Results obtained in individual laboratories may differ.
compounds on assay performance was tested. Interferences were tested
up to the listed concentrations and no impact on results was observed. Precision
Precision was determined using Elecsys reagents, samples and controls in
Endogenous substances a protocol (EP05‑A3) of the CLSI (Clinical and Laboratory Standards
Institute): 2 runs per day in duplicate each for 21 days (n = 84). The
Compound Concentration tested following results were obtained:
Bilirubin ≤ 1129 µmol/L or ≤ 66 mg/dL
cobas e 402 and cobas e 801 analyzers
Hemoglobin ≤ 1.24 mmol/L or ≤ 2000 mg/dL
Repeatabilityc) Intermediate
Intralipid ≤ 2000 mg/dL precisiond)
Biotin ≤ 410 nmol/L or ≤ 100 ng/mL Sample Mean SD CV SD CV
Rheumatoid factors ≤ 2400 IU/mL COI COI % COI %
Albumin ≤ 7.0 g/dL HSe), negative 1.12 0.015 1.3 0.017 1.5
IgG ≤ 7.0 g/dL HS, weakly positive 0.891 0.010 1.1 0.014 1.5
IgA ≤ 1.6 g/dL HS, positive 0.313 0.005 1.7 0.007 2.3
IgM ≤ 1.0 g/dL PCf) Anti-HBe 1 1.48 0.020 1.3 0.021 1.4
Criterion: Samples with a COI > 1.0: ± 20 % recovery; samples with a PC Anti-HBe 2 0.640 0.008 1.2 0.010 1.5
COI ≤ 1.0: ± 0.20 recovery.
c) Repeatability = within-run precision
Samples should not be taken from patients receiving therapy with high d) Intermediate precision = within-laboratory precision
biotin doses (i.e. > 5 mg/day) until at least 8 hours following the last biotin
e) HS = human serum
administration.
f) PC = PreciControl
Pharmaceutical substances
In vitro tests were performed on 16 commonly used pharmaceuticals. No Analytical specificity
interference with the assay was found. No cross-reactions with HAV, HCV, HIV* 1+2, HTLV**, CMV**, EBV, HSV,
In addition, the following special drugs used in hepatitis B therapy were E. coli, Toxoplasma gondii, Rubella, and Treponema pallidum were
tested. No interference with the assay was found. observed.
Measurements were performed on each of the pathogens listed above
Special drugs using ≥ 8 serum or plasma samples which were positive for antibodies to
Drug Concentration tested the above-mentioned pathogens or contained autoantibodies (SLE, AMA).
* 1 out of 16 samples was indeterminate.
mg/L
** 1 out of each 20 samples was indeterminate.
Peginterferon alfa‑2a ≤ 0.18
Clinical sensitivity
Peginterferon alfa‑2b ≤ 0.08
Samples from patients in various stages of HBV infection and from patients
Lamivudine ≤ 300 in a high-prevalence group (HBsAg and/or anti‑HBc positive) were
investigated using the Elecsys Anti‑HBe assay and various comparison
Adefovir ≤ 10

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Elecsys Anti-HBe
tests. All samples showing discrepant measurements were in the vicinity of 13 Marcellin P. Hepatitis B and hepatitis C in 2009. Liver Int
the cutoff. 2009;29(S1):1-8.
Patient Number Elecsys Anti- Anti-HBe Discrepant 14 Occupational Safety and Health Standards: Bloodborne pathogens.
(29 CFR Part 1910.1030). Fed. Register.
samples tested HBe assay comparison
positive / tests positive / 15 Directive 2000/54/EC of the European Parliament and Council of
18 September 2000 on the protection of workers from risks related to
negative negative exposure to biological agents at work.
Past HBV 192 173 / 19 154 / 38 19 For further information, please refer to the appropriate operator’s manual for
infection the analyzer concerned, the respective application sheets, the product
Chronic HBV 59 37 / 22 36 / 23 1 information and the Method Sheets of all necessary components (if
available in your country).
infection
A point (period/stop) is always used in this Method Sheet as the decimal
High 153 77 / 76 75 / 78 2 separator to mark the border between the integral and the fractional parts of
prevalence a decimal numeral. Separators for thousands are not used.
group Symbols
Clinical specificity Roche Diagnostics uses the following symbols and signs in addition to
those listed in the ISO 15223‑1 standard (for USA: see dialog.roche.com for
Samples from blood donors which had not been selected and hospitalized definition of symbols used):
patients were used to determine the specificity.
Group Number Confirmed Elecsys Specificity* Specificity** Contents of kit
tested positive Anti‑HBe react­ % % Analyzers/Instruments on which reagents can be used
ive
Reagent
Blood donors 1000 12 13 99.9 100
Calibrator
* Confirmed positive samples (i.e. confirmed by another anti‑HBe test and positive anti‑HBc and anti‑HBs find­
ings) were not considered for calculation of the % specificity.
Volume for reconstitution
** Confirmed positive samples and one sample with unclear HBV‑serology were not considered for calculation of
GTIN Global Trade Item Number
the % specificity.

204 out of 242 samples from hospitalized patients, pregnant women, and COBAS, COBAS E, ELECSYS and PRECICONTROL are trademarks of Roche. INTRALIPID is a trademark of
Fresenius Kabi AB.
dialysis patients (without symptoms of existing HBV infection) were
negative with the Elecsys Anti‑HBe assay; with a comparison test the All other product names and trademarks are the property of their respective owners.
figures were 202 out of 242. 38 samples were found to be positive by both Additions, deletions or changes are indicated by a change bar in the margin.
tests. Two samples were negative in the Elecsys Anti‑HBe assay, positive © 2021, Roche Diagnostics
in the comparison test and positive for anti‑HBc antibodies.
References
1 World Health Organization (WHO), 2015. Hepatitis B. Fact sheet
Roche Diagnostics GmbH, Sandhofer Strasse 116, D-68305 Mannheim
N°204. Available at: www.roche.com
http://www.who.int/mediacentre/factsheets/fs204/en/.
2 Kim do Y, Han KH. Epidemiology and Surveillance of Hepatocellular
Carcinoma. Liver Cancer. 2012;1(1):2-14.
3 Liang TJ. Hepatitis B: The Virus and Disease. Hepatology. 2009;49(5
Suppl):13-21.
4 Schweitzer A, Horn J, Mikolajczyk RT, et al. Estimations of worldwide
prevalence of chronic hepatitis B virus infection: a systematic review of
data published between 1965 and 2013. Lancet.
2015;386(10003):1546-1555.
5 Song Y, Bian Y, Petzold M, et al. Prevalence and Trend of Major
Transfusion-Transmissible Infections among Blood Donors in Western
China, 2005 through 2010. PLoS One. 2014 Apr 8;9(4):e94528.
6 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.
7 World Health Organization (WHO), 2009. Screening Donated Blood for
Transfusion-Transmissible Infections. Recommendations. Available at:
http://www.who.int/bloodsafety/ScreeningTTI.pdf (last access January,
2016).
8 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.
9 Liaw YF, Chu CM. Hepatitis B virus infection. Lancet
2009;373:582-592.
10 Turgeon ML. Immunology & Serology in Laboratory Medicine, 2013 5th
edition, Elsevier Health Sciences, Missouri, USA. Chapter 23.
11 Liaw YF. HBeAg seroconversion as an important end point in the
treatment of chronic hepatitis B. Hepatol Int 2009;3:425-433.
12 Negro F. Management of chronic hepatitis B: an update. Swiss Med
Wkly 2011;141:w13264.

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