Professional Documents
Culture Documents
Anti-HCV
Antibody to hepatitis C virus (anti-HCV)
REF 03290352 190 100 tests R2 Buffer (black cap), 1 bottle, 7 mL:
HEPES buffer, pH 5.0.
• Indicates analyzers on which the kit can be used
R1a Lyophilized HCV antigens, biotinylated (gray cap),
MODULAR 1 bottle for 1.2 mL solution.
Elecsys 2010 cobas e 411 cobas e 601 R2a Lyophilized HCV antigens, ruthenylated (black cap),
ANALYTICS E170
1 bottle for 1.2 mL solution.
• • • • R1b Reconstitution medium for bottle R1a (gray cap), 1 bottle, 1.4 mL:
Water, preservative.
English R2b Reconstitution medium for bottle R2a (black cap), 1 bottle, 1.4 mL:
Water, preservative.
Intended use Cal1 Negative calibrator 1 (white cap), 2 bottles of 1.3 mL each:
The Elecsys Anti-HCV assay is an in vitro diagnostic test for the qualitative Human serum, preservative.
detection of antibodies to hepatitis C virus (HCV) in human serum or plasma. Cal2 Positive calibrator 2 (black cap), 2 bottles of 1.3 mL each:
The electrochemiluminescence immunoassay “ECLIA” is intended for Human serum positive for anti-HCV Ab; preservative.
use on Elecsys and cobas e immunoassay analyzers. Non-reactive for HBsAg, anti-HIV 1/2.
b) HEPES = [4-(2-hydroxyethyl)-piperazino]-ethane sulfonic acid
Regulatory approval
The Elecsys Anti-HCV assay has been CE marked according to Directive Precautions and warnings
98/79/EC. Test performance has been established and certified by a Notified For in vitro diagnostic use.
Body according to Common Technical Specifications (2002/364/EC) for Exercise the normal precautions required for handling all laboratory reagents.
diagnostic use and for screening of blood donations. Disposal of all waste material should be in accordance with local guidelines.
Safety data sheet available for professional user on request.
Summary All human material should be considered potentially infectious.
Hepatitis C virus, first identified in 1989,1 is the most common cause All products derived from human blood are prepared exclusively from
of posttransfusion and community-acquired non-A, non-B hepatitis the blood of donors tested individually and shown to be free from
worldwide. Infection with HCV frequently leads to chronic hepatitis and HBsAg and antibodies to HCV (Cal1 only) and HIV.
cirrhosis, and is associated with the development of hepatocellullar The testing methods applied were FDA-approved or cleared in compliance
carcinoma.2 Common extrahepatic manifestations comprise mixed with the European Directive 98/79/EC, Annex II, List A.
cryoglobulinemia and other rheumatic diseases.3 The serum containing anti-HCV (Cal2) was inactivated using
Hepatitis C virus is an enveloped, positive-sense single-stranded RNA virus β-propiolactone and UV-radiation.
which has been classified as an own genus in the family of Flaviviridae. The However, as no inactivation or testing method can rule out the potential
genome consists of ~9.5 kb encoding for a 3000 amino acid polypeptide of risk of infection with absolute certainty, the material should be treated just
structural and non-structural domains.4 Like other RNA viruses, the HCV as carefully as a patient specimen. In the event of exposure the directives
genome exhibits substantial heterogeneity as a result of mutations that occur of the responsible health authorities should be followed.11,12
during viral replication. Worldwide, at least 11 genetically distinct genotypes The reagents may not be used after the stated expiration date.
and multiple subtypes and virus variants have been described.5 Infection with Avoid the formation of foam with all reagents and sample types
specific genotypes can affect disease severity and treatment response.6,7 (specimens, calibrators, and controls).
Hepatitis C is primarily transmitted through contaminated blood and blood The Elecsys Anti-HCV assay has a high dilution sensitivity. Avoid any
products and to a lower extent by human body secretions.8 sample cross-contamination during sample pre-analytics.
Anti-HCV antibody tests are used alone or in combination with other tests
(e.g. HCV-RNA) to detect an infection with hepatitis C virus and to identify Reagent handling
blood and blood products of individuals infected with HCV. The reagents R1 and R2 are not ready for use and have to be
The Elecsys Anti-HCV assay is a third-generation test.9,10 The assay prepared. See “Preparation of working solutions” section for further
uses peptides and recombinant antigens representing core, NS3 and instructions. The reagents M, Cal1 and Cal2 are ready for use and
NS4 proteins for the determination of anti-HCV antibodies. are supplied in bottles compatible with the system.
Elecsys 2010 and cobas e 411 analyzers: The calibrators Cal1 and
Test principle Cal2 should only be left on the analyzer during calibration at 20-25 °C.
Sandwich principle. Total duration of assay: 18 minutes. After use, close the bottles as soon as possible and store at 2-8 °C.
• 1st incubation: 40 µL of sample, 60 µL of a reagent containing biotinylated Ensure that no calibration solution is trapped in the opened snap-cap.
HCV antigens and 60 µL of a reagent containing HCV antigens labeled Because of possible evaporation effects, not more than 5 calibration
with a ruthenium complexa react to form a sandwich complex. procedures per calibrator bottle set should be performed.
• 2nd incubation: After addition of streptavidin-coated microparticles, MODULAR ANALYTICS E170 and cobas e 601 analyzers: Unless
the complex becomes bound to the solid phase via interaction the entire volume is necessary for calibration on the analyzer, transfer
of biotin and streptavidin. aliquots of the ready-for-use calibrators into empty snap-cap bottles
• The reaction mixture is aspirated into the measuring cell where the (CalSet Vials). Attach the supplied labels to these additional bottles.
microparticles are magnetically captured onto the surface of the Store the aliquots for later use at 2-8 °C.
electrode. Unbound substances are then removed with ProCell. Perform only one calibration procedure per aliquot.
Application of a voltage to the electrode then induces chemiluminescent All information required for correct operation is read in via the
emission which is measured by a photomultiplier. respective reagent barcodes.
• Results are determined automatically by the Elecsys software by
comparing the electrochemiluminescence signal obtained from the Preparation of working solutions
sample with the cutoff value obtained by anti-HCV calibration. The reagents R1 and R2 are not ready for use and have to be
prepared by adding the reconstituted antigens.
a) Tris(2,2’-bipyridyl)ruthenium(II)-complex (Ru(bpy)2+
3 )
For the reconstitution of the lyophilized antigens proceed as follows:
Reagents - working solutions A. Using adapters
M Streptavidin-coated microparticles (transparent cap), 1 bottle, 6.5 mL: 1a. Connect bottle R1a (lyophilized biotinylated antigens; gray cap)
Streptavidin-coated microparticles 0.72 mg/mL; preservative. with bottle R1b (reconstitution medium for bottle R1a; gray cap)
R1 Buffer (gray cap), 1 bottle, 7 mL: using one of the adapters. Transfer the volume of reconstitution
HEPESb buffer, pH 5.0. medium. Avoid the formation of foam!
Anti-HCV
Antibody to hepatitis C virus (anti-HCV)
• REF 12102137001, AssayTip/AssayCup Combimagazine M, 48 magazines Calculation
x 84 reaction vessels or pipette tips, waste bags The analyzer automatically calculates the cutoff based on the
• REF 03023150001, WasteLiner, waste bags measurement of Cal1 and Cal2.
• REF 03027651001, SysClean Adapter M The result of a sample is given either as reactive or non-reactive as well
as in the form of a cutoff-index (signal sample/cutoff).
Accessories for all analyzers:
Interpretation of the results:
• REF 11298500316, Elecsys SysClean, 5 x 100 mL system cleaning solution
Samples with a cutoff-index < 0.9 are non reactive in the
Assay Elecsys Anti-HCV assay.
For optimum performance of the assay follow the directions given in Samples with a cutoff-index ≥ 1.0 are reactive in the Elecsys Anti-HCV assay.
this document for the analyzer concerned. Refer to the appropriate All samples reactive in the initial test must be redetermined in
operator’s manual for analyzer-specific assay instructions. duplicate using the Elecsys Anti-HCV assay.
Resuspension of the microparticles takes place automatically before use. Samples having a cutoff-index between ≥ 0.9 and < 1.0 are considered to
Read in the test-specific parameters via the reagent barcode. If in exceptional be in the gray zone. All initially reactive samples in the gray zone have to
cases the barcode cannot be read, enter the 15-digit sequence of numbers. be retested in duplicate with the Elecsys Anti-HCV assay.
MODULAR ANALYTICS E170 and cobas e 601 analyzers: If no reactivity is found in both cases, the sample is negative for anti-HCV.
PreClean M solution is necessary. If the result from either of the two measurements is reactive or borderline
MODULAR ANALYTICS E170, Elecsys 2010 and cobas e analyzers: Bring then the sample is repeatedly reactive. Repeatedly reactive samples
the cooled reagents to approx. 20 °C and place on the reagent disk (20 °C) of must be investigated by supplemental methods (e.g. immunoblot or
the analyzer. Avoid the formation of foam. The system automatically regulates detection of HCV RNA). If one or both measurements remain borderline
the temperature of the reagents and the opening/closing of the bottles. the analysis of a follow-up sample is recommended.
Place the calibrators Cal1 and Cal2 in the sample zone of the analyzer. Limitations - interference
Only keep open during calibration. All information necessary for calibration The assay is unaffected by icterus (bilirubin < 855 µmol/L or < 50 mg/dL),
is encoded on the barcoded bottle label and is read in automatically. After hemolysis (Hb < 1.09 mmol/L or < 1.75 g/dL), lipemia (Intralipid
calibration has been performed, store Cal1 and Cal2 at 2-8 °C or discard < 2100 mg/dL) and biotin < 50 ng/mL.
(MODULAR ANALYTICS E170 and cobas e 601 analyzers). Criterion: Recovery of positive samples within ± 20 % of initial value,
Calibration cutoff-index for negative samples ≤ 0.5.
Calibration frequency: In patients receiving therapy with high biotin doses (i.e. > 5 mg/day), no sample
Every Elecsys Anti-HCV rackpack must be calibrated using should be taken until at least 8 hours after the last biotin administration.
Elecsys Anti-HCV Cal1 and Cal2. Lot calibrations are not allowed No interference was observed from rheumatoid factors up to
for the Elecsys Anti-HCV assay. a concentration of 1000 IU/mL.
Renewed calibration for each Elecsys Anti-HCV rackpack No false negative result due to high-dose hook effect was found
is recommended as follows: with the Elecsys Anti-HCV assay.
In vitro tests were performed on 18 commonly used pharmaceuticals and 2
• after 7 days (when the same reagent kit is alternately used
drugs used in HCV therapy. No interference with the assay was found.
on the analyzer and refrigerated)
In rare cases, interference due to extremely high titers of antibodies
• after 12-24 hours if the preparation of working solutions is performed to streptavidin and ruthenium can occur.
according to 6b. (using adapters) or 5b. (by pipetting) For diagnostic purposes, the results should always be assessed in conjunction
• as required: e.g. quality control findings outside the specified limits with the patient’s medical history, clinical examination and other findings.
• more frequently when this is required by pertinent regulations Due to a long time period from infection to seroconversion, negative anti-HCV
Range for electrochemiluminescence signals (counts) for the calibrators: test results may occur during early infection. If acute hepatitis C infection is
Negative calibrator (Cal1): 350-1200 (Elecsys 2010 and cobas e 411 analyzers) suspected, measuring of HCV RNA by reverse transcriptase polymerase chain
or 350-1600 (MODULAR ANALYTICS E170 and cobas e 601 analyzers). reaction (RT-PCR e.g. by Cobas Amplicor) may give evidence of HCV infection.
Positive calibrator (Cal2): 10000-38000 (Elecsys 2010 and cobas e The detection of anti-HCV antibodies indicates a present or past infection with
411 analyzers) or 10000-60000 (MODULAR ANALYTICS E170 hepatitis C virus, but does not differentiate between acute, chronic or resolved
and cobas e 601 analyzers). infection. It is recognized within the scientific community that presently
No internationally accepted standard for anti-HCV exists. available methods for anti-HCV detection are not sensitive enough to detect all
potentially infectious units of blood or possible cases of HCV infection. The
Quality control
antibody concentration may be beneath the detection limit of this assay or the
For quality control, use Elecsys PreciControl Anti-HCV 1 and 2.
patient´s antibodies do not react with the antigens used in this test. In addition,
The controls 1 and 2 should be run as single determinations at least once every
non-specific results cannot be ruled out with the Elecsys Anti-HCV assay.
24 hours when the test is in use, once per reagent kit, and after every calibration.
The control intervals and limits should be adapted to each laboratory’s Specific performance data
individual requirements. Values obtained should fall within the defined limits. Representative performance data on the analyzers are given below.
Each laboratory should establish corrective measures to be taken Results obtained in individual laboratories may differ.
if values fall outside the limits.
If necessary, repeat the measurement of the samples concerned. Precision
Follow the applicable government regulations and local guidelines Precision was determined using Elecsys reagents, human sera, and
for quality control. controls (repeatability n = 21, intermediate precision n = 10); total
precision on MODULAR ANALYTICS E170 analyzer was determined
Note:
in a modified protocol (EP5-A) of the CLSI (Clinical and Laboratory
For technical reasons re-assigned target values valid for a specific reagent
Standards Institute): 6 times daily for 10 days (n = 60).
and control lot combination only, must be entered manually. Therefore,
The following results were obtained by preparation of the working solutions via
always consider the value sheet included in the rackpack or PreciControl
adapters. Comparable results were obtained when using the pipetting method.
kit to make sure that the correct target values are used.
When a new reagent or control lot is used, the analyzer will use the
original values encoded in the control barcodes.