Anti-HCV

After incubation, anti-human IgG monoclonal antibody

conjugate binds to anti-HCV antibodies captured on
Antibody to Hepatitis C Virus（CLIA） microparticles. Microparticles are magnetically captured
while other unbound substances are removed by
Order Information washing.
Catalog No. Package Size In the third step, the substrate solution is added into the
Anti-HCV111 2×50 tests reaction cuvette. It is catalyzed by alkaline phosphatase
Anti-HCV112 2×100 tests in the immune complex captured on the microparticles.
The resulting chemiluminescent reaction is measured as
Intended Use relative light units (RLUs) by a photomultiplier built
The CL-series Anti-HCV assay is a Chemiluminescent
inside the system. A direct relationship exists between
Immunoassay (CLIA) for the qualitative determination
the amount of anti-HCV antibodies in the sample and the
of antibody to hepatitis C virus (Anti-HCV) in human
RLUs generated during the reaction.
serum or plasma.
The presence or absence of anti-HCV antibodies in the

Summary sample is determined by comparing the

Hepatitis C virus (HCV) is an enveloped, positive-sense chemiluminescent signal of the reaction to the cutoff
single-stranded RNA virus which has been classified as signal determined from system calibration. A cutoff
an own genus in the family of Flaviviridae. It is the most index (COI) is calculated by Sample RLU/Cutoff RLU.
1-3
common cause of bloodborne as well as
Reagents Components
community-acquired non-A, non-B hepatitis worldwide4.
The reagent kit is composed of four components: Ra, Rb,
The HCV genome consists of ~9.5 kb encoding for a
Rc, and Rd. The component cannot be exchanged, and
3000 amino acid polypeptide of structural and
the detailed information of each component is listed
non-structural domains5. Like other RNA viruses, and
below：
exhibits substantial heterogeneity as a result of
Paramagnetic microparticles coated with
mutations that occur during viral replication. The
Ra recombinant HCV antigens in MES buffer with
presence of anti-HCV indicates that an individual may
preservative.
have been infected with HCV, may harbor infectious HCV,
Alkaline phosphatase labeled anti-human IgG
and/or may be capable of transmitting HCV infection6.
Rb monoclonal antibody conjugate in MES buffer with
Anti-HCV antibody tests are used alone or in
preservative.
combination with other tests (e.g. HCV-RNA) to detect
Rc Sample diluents in HEPES buffer with preservative.
an infection with hepatitis C virus and to identify blood
Rd MES buffer with preservative.
and blood products of individuals infected with HCV.
The position of each component is shown in the figure
Assay Principles below (front view on the left and top view on the right):
The CL-series Ant-HCV assay is a two-step sandwich
assay to determine the level of antibody to hepatitis C
virus.
In the first step, sample, sample treatment solution,
paramagnetic microparticles coated with recombinant
HCV antigens are added into a reaction cuvette. After
incubation, antibodies to HCV present in the sample bind
Storage Stability
to HCV antigens coated on microparticles. Afterwards,
The unopened Anti-HCV (CLIA) reagent is stable up to
microparticles are magnetically captured while other
one full year when stored at 2-8 0C, the actual expiration
unbound substances are removed by washing.
date is stated on the label.
In the second step, diluent solution and alkaline
The Anti-HCV (CLIA) reagent can be stored onboard at
phosphatase labeled anti-human IgG monoclonal
2-8 0C and used for a maximum of 28 days after opening
antibody conjugate are added into the reaction cuvette.
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for use. use grossly hemolyzed specimens. Do not use
heat-inactivated specimens.
Reagent Preparation
Specimens should be tested as soon as possible after
Ra: Ready to use
sample collection. If testing is not completed within 8
Rb: Ready to use
hours, specimens should be stored at 2-8°C or colder.
Rc: Ready to use
Specimens would be stable for 7 days at 2-8°C, 3
Rd: Ready to use
months at -20°C. Avoid more than five freeze cycles.
Materials required but not provided
Assay Procedure
Mindray CL-series Chemiluminescence Immunoassay
For optimal performance of this assay, operators should
Analyzer.
read the related system operation manual carefully to get
Cat. No. Anti-HCV211: Anti-HCV Calibrators, 1×2.0mL
sufficient information, such as operation instructions,
for each level of calibrator C0 and C1.
sample preservation and management, safety
Cat. No. Anti-HCVN311: Anti-HCV Negative Control
precautions, and maintenance. Prepare all required
3×2.0mL.
materials for the assay as well.
Cat. No. Anti-HCVN312: Anti-HCV Negative Control,
Before loading the Anti-HCV reagent kit on the
6×2.0mL.
instrument for the first time, invert unopened reagent
Cat. No. Anti-HCVP311: Anti-HCV Positive Control,
bottle gently for at least 30 times to resuspend the
3×2.0mL.
microparticles, which have settled during shipment or
Cat. No. Anti-HCVP312: Anti-HCV Positive Control,
storage. Visually inspect the bottle to ensure the
6×2.0mL.
microparticles have been resuspended. If the
Cat. No. CS511: Substrate Solution, 4×115mL.
microparticles remain adhered to the bottle, continue
Cat. No. WB411: Wash Buffer.
inverting until the microparticles have been completely
Reaction Vessel.
resuspended. If the microparticles cannot be
Instrument System homogenized, it is recommended not to use this bottle of
Mindray CL-series Chemiluminescence Immunoassay reagent. Contact Mindray Customer Service for help. Do
Analyzer. not invert opened reagent bottle.
This assay requires 20 μL of sample volume for a single
Specimen Collection and Preparation
test. This volume does not include the dead volume of
Human serum and plasma collected sodium heparin and
the sample container. Additional volume is required
lithium heparin are recommended for this assay.
when performing additional tests from the same sample.
Collect all blood samples observing routine precautions
Operators should refer to the system operation manual
for venipuncture. Follow blood collection tube
and specific requirement of the assay to determine the
manufacturer’s recommendations for centrifugation.
minimum sample volume.
Centrifuge the specimens after clot formation is
complete. Some specimens, especially those from Calibration
patients receiving anticoagulant therapy may exhibit The calibration information is stored in the barcode
increased clotting time. Please ensure that residual attached in the reagent and calibrator pack. When
fibrin and cellular matter have been removed prior to performing the calibration, scan the information from
analysis. the barcodes into the system first, and then test the
For optimal results, inspect all samples for bubbles. calibrators at two levels. Calibration is required before
Remove bubbles with a pipette tip prior to analysis. any Anti-HCV test. Recalibration is recommended every
Specimens must be mixed thoroughly after thawing. 4 weeks, or when a new reagent lot is used, or the
Thawed samples should be centrifuged prior to use. If quality controls are out of specified range. For detailed
the sample was covered with lipid layer after instruction of calibration, refer to the system operation
centrifugation, the sample should be transferred to a manual.
clean tube before testing. Do not transfer the lipid layer.
Quality Control
Handle carefully to prevent cross contamination. Do not
For verifying the reliability of assay system, quality
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controls should be run at least once every 24 hours, after and clinical history, etc. to make clinical decisions.
a new reagent pack is loaded, or after every calibration.
Performance Characteristics
The recommended quality controls for this assay are
Mindray Anti-HCV Negative Control and Anti-HCV Positive Sensitivity
Control. 400 specimens have been claimed to be anti-HCV
Quality control results should be within the acceptable positive by reference methods. Mindray Anti- HCV assay
ranges. If a control is out of its specified range, the on CL-2000i system has detected 399 samples as
associated test results are invalid and the samples must positive and 1 samples as negative. The positive
be retested. Recalibration may be required. Refer to the coincidence rate is 99.75%.
system operation manual to check up the system. If the
Precision
quality control results are still out of the specified ranges,
The CL-series Anti-HCV assay is designed to have a
please contact Mindray Customer Service for help.
precision of ≤10% (within-device CV). Precision was
Calculation determined by following National Committee for Clinical
The CL-series analyzer automatically calculates Cutoff Laboratory Standards (NCCLS) Protocol EP5-A27. Two
RLU using the mean RLU from three replicates of levels of quality controls were tested in duplicate in two
calibrator C0 and two replicates of calibrator C1, and separate runs per day, for a total of 20 days, using a
stores the results. Then the analyzer calculates single lot of reagents and a single calibration curve. The
Anti-HCV result based on the cutoff RLU by the following precision data are summarized in the table below.
calculation.
Mean Anti-HCV Within- Between- Within-
Cutoff RLU = [(Mean RLU of C1 – Mean RLU of C0) x Sample
(COI) Run CV Run CV Device CV
Calibration Coefficient] + Mean RLU of C0.
Negative Control 0.21 2.09% 5.34% 8.09%
Calibration Coefficient is specific for each combination of
Positive Control 3.93 2.48% 1.69% 3.34%
reagent kit lot and calibrator lot.
COI = Sample RLU/Cutoff RLU. Specificity
Hemoglobin up to 500 mg/dL, bilirubin up to 20 mg/dL,
Interpretation of Results
triglycerides up to 3000 mg/dL, and total protein up to
Samples with a COI<1.00 are Non-reactive in the
10 g/dL will not interfere with the CL-series Anti-TP assay.
Mindray Anti-HCV assay. These samples do not need
With these substances at indicated concentration , the
further testing.
difference is < 0.1 COI on negative specimens and ＜
Samples with a COI≥1.00 are Initially Reactive in the
10% COI on positive samples .
Mindray Anti-HCV assay. All Initially Reactive samples
No obvious interference was observed from rheumatoid
should be transferred to a centrifuge tube and
factor (up to 1000 IU/mL) or antinuclear antibody.
centrifuged at≥10,000 RCF (Relative Centrifugal Force)
Mindray Anti-HCV assay was evaluated for potential
for 10 minutes, and retested in duplicate. If COI<1.00 in
cross-reactive substances from individuals with medical
both retests, the sample is considered as Non-reactive
conditions unrelated to HCV infection. At least 5 samples
for Anti-HCV. If COI>1.00 in either of the retests, the
containing each of the potentially interfering pathogens of
sample is Repeatedly Reactive for Anti-HCV.
the following (HAV, HEV, HBV, CMV, HSV, HIV-1/2, Toxo,
Repeatedly reactive samples should be investigated by
RV, TP, EBV, E.coli）are tested by Mindray Anti-HCV and a
supplemental tests such as other HCV specific
reference reagent kit. All results are consistent between
immunoassays and immunoblot assays or a combination
Mindray assay and the reference assay.
thereof and/or NAT tests.
623 specimens have been claimed as Anti-HCV negative
Limitation of Measurement by reference methods. Mindray Anti-HCV assay on
The Anti-HCV result of a given specimen can vary, in CL-2000i system detected 621 samples as negative and
assays from different manufacturers, due to differences 2 samples as positive. The negative coincidence rate is
in assay methods, calibration, and reagent specificity. 99.68%.
The assay results should be used in conjunction with
other data, such as symptoms, results of other tests,
Warnings and Precautions

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 1. For in vitro diagnostic use only. 4. Alter MJ, et al. Risk Factors for Acute Non-A, Non-B
2. Follow all the rules in handling laboratory Hepatitis in the United States and Association with
reagents and take necessary safety precautions. Hepatitis C Virus Infection. JAMA 1990;264:2231-5.
3. Due to the differences in methodology and 5. Major ME, Feinstone SM. The Molecular Virology of
antibody specificity, test results of the same sample may Hepatitis C. Hepatology 1997;25(6):1528-1538.
be different when using reagent kits from different 6. Choo QL, et al. Hepatitis C Virus: The Major
manufacturers on Mindray system, or using Mindray Causative Agent of Viral Non-A, Non-B Hepatitis. Br
reagent kits on other systems. Med J 1990;46:423-41.
4. Do not use reagent kits beyond the expiration 7. CLSI. EP5-A2: Vol. 24, No. 25, Evaluation of
date. Precision Performance of Quantitative Measurement
5. Do not use reagents mixed from different reagent Method; Approved Guideline –Second Edition.
lots.
6. Always keep the reagent pack in the upright © 2016 Shenzhen Mindray Bio-Medical Electronics Co.,
position to ensure no microparticle has been lost prior to Ltd.
use. All rights Reserved
7. Reagent pack opened for more than 28 days is not Manufacturer: Shenzhen Mindray Bio-Medical
recommended for use. Electronics Co., Ltd.
8. Reliability of assay results cannot be guaranteed if Address: Mindray building, Keji 12th Road South,
the instructions in this package insert are not followed. Hi-tech Industrial Park, Nanshan, Shenzhen, 518057
9. All the specimen and reaction wastes should be P.R.China
considered potentially biohazard. Specimens and E-mail Address: [email protected]
reaction wastes should be handled in accordance with Tel: +86-755-26582888
the local regulations and guidelines. Fax: +86-755-26582680
10. The Material Safety Data Sheet (MSDS) is
available upon request. EC-Representative: Shanghai International Holding
Corp. GmbH (Europe)
Graphical Symbols
Address: Eiffestraβe 80, Hamburg 20537, Germany
Tel: 0049-40-2513175
Fax: 0049-40-255726

Bibliography
1. Alter HJ, et al. Detection of Antibody to Hepatitis C
Virus in Prospectively Followed Transfusion
Recipients with Acute and Chronic Non-A, Non-B
Hepatitis. N Engl J Med 1989;321:1494-500.
2. Esteban JI, et al. Hepatitis C Virus Antibodies Among
Risk Groups in Spain. Lancet 1989;ii:294-7.
3. Aach RD, et al. Hepatitis C Virus Infectionin
Post-Transfusion Hepatitis: An Analysis with First-
and Second-Generation Assays. N Engl J Med
1991;325:1325-9.

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