HIV

assay also target HIV p24 antigen in serum or plasma for

identification of the recently infected individuals.14, 15
Antigen and Antibodies to Human Mindray HIV assay belongs to the 4th generation HIV
Immunodeficiency Virus（CLIA） assay which detects both HIV-1 (including Group M & O)
and HIV-2 antibodies, as well as HIV p24 antigen prior to
Order Information seroconversion, thereby shortening the detection
Catalog No. Package Size window and improving detection sensitivity of early HIV
HIV111 2×50 tests infection.
HIV112 2×100 tests
Assay Principles
Intended Use The CL-series HIV assay is a two-site sandwich assay to
The CL-series HIV assay is a Chemiluminescent determine the level of HIV p24 antigen and the
Immunoassay for the simultaneous qualitative antibodies to HIV-1 and/or HIV-2.
determination of HIV p24 antigen, and antibodies to In the first step, sample, sample treatment solution,
HIV-1 and/or HIV-2 in human serum or plasma. paramagnetic microparticles coated with mouse
anti-HIV p24 mAb, and microparticles coated with
Summary specific HIV-1 and HIV-2 antigens are added into a
Human Immunodeficiency Virus (HIV）is the etiologic
reaction vessel. After incubation, HIV p24 antigen in the
agent causing acquired immunodeficiency syndrome
sample will bind specifically to the monoclonal antibody
(AIDS), which belongs to retrovirus，classified in the
coated on microparticles, while HIV-1 and HIV-2
category of Lentivirus, primate immunodeficiency
antibodies in the sample will bind to HIV-1 or HIV-2
subgenus. HIV is transmitted by sexual contact, exposure
antigen coated microparticles. Afterwards,
to blood or blood products, or prenatal infection of a fetus
microparticles are magnetically captured while other
or a newborn.1
unbound substances are removed by washing.
There are two types of human immunodeficiency viruses,
In the third step, diluent solution, ALP labeled anti-HIV
HIV-1 and HIV-2, both composed of multiple genetic
p24 monoclonal antibody, and ALP labeled HIV-1 and
subtypes2, 3. The prevalent subtypes in different areas
HIV-2 antigens are added to the reaction vessel. After
could vary. HIV-1 is classified into three main groups:
incubation, ALP labeled anti-HIV p24 monoclonal
Group M (major), Group O (outlier), and Group N (Non-M
antibody, ALP labeled HIV-1 and HIV-2 antigen will form
and Non-O).4-6 Among HIV-1 Group M there are nine
sandwich with microparticle captured HIV p24 antigen,
subtypes (A, B, C, D, F, G, H, J & K) and circulating
HIV-1 or HIV-2 antibody respectively. Afterwards,
recombinant forms (CRFs). HIV-1 is the most widely
microparticles are magnetically captured while other
transmitted HIV virus.4 HIV-1 Group O and Group N were
unbound substances are removed by washing.
discovered in Cameron, which are rare and only endemic
In the third step, the substrate solution is added to the
to West Central Africa. However, Group O infections have
reaction vessel. The substrate is catalyzed by ALP in the
5, 6
been identified in other areas too. HIV-2 is classified
immune complex captured on the microparticles, The
into genetic subtypes A-H, among which A and B are the
resulting chemiluminescent reaction is measured as
most endemic subtypes.7
relative light units (RLUs) by a photomultiplier built into
When HIV infects human being, the immune system of
the system. A direct relationship exists between the
human body will respond by generating a variety of
amount of p24 antigen, and antibody to HIV-1 or HIV-2
antibodies to HIV virus proteins. The antibodies against
or both in the sample and the RLUs generated during the
trans-membrane protein (TMP) of the virus are among the
reaction. The presence or absence of HIV p24 antigen,
first to appear at seroconversion of HIV infected
HIV-1 or HIV-2 antibodies in the sample is determined
individuals. In addition, the anti-TMP response remains
by comparing the chemiluminescent signal in the
relatively strong throughout the course of the disease.
reaction to the cutoff signal determined from calibration.
Therefore, anti-TMP antibodies are good targets for HIV
A cutoff index (COI) is calculated by Sample RLU/Cutoff
assay.8-13 In addition to antibody, the 4th generation HIV
RLU.
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Reagent Components Control:6×2.0mL);
The reagent kit is composed of four components: Ra, Rb, Cat. No. HIVP312: HIV Ag/Ab Positive Control, 6×2.0mL
Rc, and Rd. The component cannot be exchanged, and (HIV Ag Positive Control:3×2.0mL, HIV Ab Positive
the detailed information of each component is listed Control:3×2.0mL);
below： Cat. No. CS511: Substrate Solution, 4×115mL;
Paramagnetic microparticles coated with Cat. No. WB411: Wash Buffer;
HIV-1/2 specific antigens and HIV p24 Reaction Vessel
Ra
monoclonal antibody (mouse IgG) in HEPES
Instrument System
buffer with preservative.
Mindray CL-series Chemiluminescence Immunoassay
ALP labeled HIV-1/2 specific antigens and HIV
Analyzer.
Rb p24 monoclonal antibody (mouse IgG) in
TRIS buffer with preservative. Specimen Collection and Preparation
Sample diluents in TRIS buffer with Human serum is recommended for this assay. Centrifuge
Rc
preservative. the specimens at 3500 rpm for a minimum of 10 minutes
Rd Blockers in TRIS buffer with preservative. after clot formation is complete. Specimens should be

The position of each component is shown in the figure tested as soon as possible after sample collection and
below（front view on the left and top view on the right）: pre-analytical treatment. Test the specimens within two
hours after centrifugation. If testing is not completed
within 12 hours, transfer the supernatant into tubes for
storage. Specimens should be tightly capped and
refrigerated at 2-8°C. If testing will be delayed for more
than 7 days, specimens should be frozen at -20°C or
below.
Avoid repeated freeze and thaw cycles, which may cause
Storage and Stability sample deterioration. Specimen can be used after a
The unopened HIV reagent kit is stable up to the
maximum of five cycles of freeze and thaw.
expiration date as indicated on the label when stored at
Do not use specimen with the following conditions:
0
2-8 C. The actual expiration date is stated on the
 heat inactivated
label.
 grossly hemolyzed
The HIV reagent kit can be stored onboard at 2-8 0C and
 obvious microbial contamination
used for a maximum of 28 days after opening for use.
 visible fibrin or other debris

Reagent Preparation
Assay Procedure
Ra: Ready to use
For optimal performance of this assay, operators should
Rb: Ready to use
read the related system operation manual carefully to get
Rc: Ready to use
sufficient information such as operation instructions,
Rd: Ready to use
sample preservation and management, safety

Materials Required but not Provided precautions, and maintenance. Prepare all required

Mindray CL-series Chemiluminescence Immunoassay materials for the assay as well.

Analyzer. Before loading the HIV reagent kit on the instrument for

Cat. No. HIV211: HIV Calibrators, 1×2.0mL for each the first time, invert unopened reagent bottle gently for
level of calibrator C0 and C1； at least 30 times to resuspend the microparticles, which

Cat. No. HIVN311: HIV Negative Control 3×2.0mL; have settled during shipment or storage. Visually inspect

Cat. No. HIVN312: HIV Negative Control, 6×2.0mL; the bottle to ensure the microparticles have been well
Cat. No. HIVP311: HIV Ag/Ab Positive Control, mixed. If the microparticles remain adhered to the bottle,

12×2.0mL continue inverting until the microparticles have been

(HIV Ag Positive Control:6×2.0mL, HIV Ab Positive completely mixed. If the microparticles cannot be
2016-08 V5.0 English HIV Page 2 of 5 P/N: 046-003271-00 (5.0)
homogenized, it is recommended not to use this bottle of COI = Sample RLU/Cutoff RLU
reagent. Contact Mindray Customer Service for help. Do
Interpretation of Results
not invert opened reagent bottle.
Samples with a COI<1.00 are Non-reactive in the
This assay requires 100 μL of sample volume for a
Mindray HIV assay. These samples are considered as
single test. This volume does not include the dead
negative for HIV, and do not need further testing.
volume of the sample container. Additional volume is
Samples with a COI I≥1.00 are considered Initially
required when performing additional tests from the
Reactive.
same sample. Operators should refer to the system
All Initially Reactive specimens must be transferred to a
operation manual and specific requirement of the assay
centrifuge tube and centrifuged at≥10,000 RCF
to determine the minimum sample volume.
(Relative Centrifugal Force) for 10 minutes, and then
Calibration retested in duplicate.
The calibration information is stored in the barcode If COI is <1.00 in both retests, this sample is considered
attached in the reagent and calibrator pack. When as negative for HIV p24 antigen and HIV-1/2 antibodies.
performing the calibration, scan the information from
If COI is ≥ 1.00 in any of the retests, the sample is
the barcodes into the system first, and then test the
considered as Repeatedly Reactive, and is positive for
calibrators at two levels. Calibration is required before
either HIV p24 antigen, or HIV-1/2 antibodies, or
any HIV test. Recalibration is recommended every 4
positive for both antigen and antibodies.
weeks, or when a new reagent lot is used, or when the
Repeatedly Reactive samples must be confirmed
quality controls are out of specified ranges. For detailed
according to the recommended confirmatory procedure
instruction of calibration, refer to the system operation
by regulatory authority, such as Western Blot and HIV
manual.
RNA tests.
Control
Limitation of Measurement
It is recommended that quality controls should be run
The HIV result of a given specimen can vary, depending
once every 24 hours if the tests are in use, or after every
on the assays from different manufacturers, which have
calibration. The quality control frequency should be
differences in assay methods, calibration, and reagent
adapted to each laboratory’s individual requirements.
specificity. The assay results should be used in
Two controls for this assay are Mindray HIV Negative
conjunction with other data, such as symptoms, results
Control and HIV Positive Control.
of other tests, and clinical history, etc. to make clinical
Quality control results should be within the acceptable
decisions.
ranges. If a control is out of its specified range, the
associated test results are invalid and the samples must Performance Characteristics
be retested. Recalibration may be required. Refer to the
Sensitivity
system operation manual to check up the system. If the
The sensitivity for HIV p24 at the cutoff is <2 IU/mL. The
quality control results are still out of the specified ranges,
measurement accuracy is traceable to WHO
please contact Mindray Customer Service for help.
International Standard HIV p24 Antigen (NIBSC code:
Calculation 90/636).
The CL-series analyzer calculates the mean RLU signals 505 specimens have been clinically diagnosed with HIV
of both calibrator C1 and C0. infection and classified as disease status. Among them,
The CL-series analyzer calculates HIV result based on the five specimens are HIV-O antibody positive, 30 are
ratio of the sample RLU to the cutoff RLU (Cutoff Index, HIV-2 antibody positive, and 23 are HIV p24 antigen
COI) for each specimen and control. positive. HIV assay on Mindray CL-2000i system
Cutoff RLU = [(Mean RLU of C1 – Mean RLU of C0) x detected 505 specimens as positive, with the positive
Calibration Coefficient] + Mean RLU of C0 coincidence rate at 100%..
Calibration Coefficient is specific for each reagent and
Precision
calibrator lot.
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The CL-series HIV assay is designed to have a precision 4. Do not use reagent kits beyond the expiration
of ≤10% (within-device CV). Precision was determined date.
by following National Committee for Clinical Laboratory 5. Do not use reagents mixed from different reagent
14
Standards (NCCLS) Protocol EP5-A2 . Three levels of lots.
quality controls were tested in duplicate in two separate 6. Always keep the reagent pack in the upright
runs per day, for a total of 20 days, using a single lot of position to ensure no microparticle has been lost prior to
reagents and a single calibration curve. The precision use.
data are summarized in the table below. 7. Reagent pack opened for more than 28 days is not
Average Within- Between- Within- recommended for use.
Sample
(COI) Run CV Run CV Device CV 8. Reliability of assay results cannot be guaranteed if
Negative Control 0.14 2.42% 3.44% 6.46% the instructions in this package insert are not followed.
Positive Control 1 6.95 1.15% 2.98% 3.40% 9. All the specimen and reaction wastes should be
Positive Control 2 5.95 1.18% 3.02% 3.66% considered potentially biohazard. Specimens and
reaction wastes should be handled in accordance with
Specificity
the local regulations and guidelines.
Hemoglobin up to 500 mg/dL, bilirubin up to 20 mg/dL,
10. The Material Safety Data Sheet (MSDS) is
triglycerides up to 3000 mg/dL, total protein up to 10
available upon request.
g/dL, rheumatoid factor(RF) up to 1500 IU/mL,
anti-DNA antibody (ANA) up to 400 U/L, and mouse Graphical Symbols
anti-human antibody（HAMA）will not interfere with the
HIV assay. These substances show less than 10%
interference at indicated concentrations.
Mindray HIV kit was evaluated for potential
cross-reactive substances from individuals with medical
conditions unrelated to HIV infection. At least 5 samples
containing each of the potentially interfering substance
of the following (HAV, Anti-HBs, HCV, HEV, HGV,
Anti-HTLV-Ⅰ/Ⅱ, HBsAg, TP, EBV, RV, CMV, HSV-1, Toxo,
Bibliography
Rubella, E.coli, Yeast, and Lentivirus ） are tested by
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Mindray HIV and a reference reagent kit. All results are
Comprehensive Guide to Technical, Medical, Social,
consistent between Mindray assay and the reference
Legal, and Management Issues. 2nd ed. New York:
assay.
NY Springer-Verlag; 1994.
1021 specimens have been claimed negative by
2. Barre-Sinoussi F, Chermann JC, Rey F, et al.
reference methods. HIV assay on Mindray CL-2000i
Isolation of a T-lymphotropicretrovirus from a
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3. Clavel F, Guetard D, Brun-Vezinet F et al. Isolation

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2. Follow all the rules in handling laboratory 4. Peeters, M. Recombinant HIV sequences: their role

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3. Due to the differences in methodology and et al, editors. HIV Sequence Compendium 2000. Los

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be different when using reagent kits from different 2000:54-73. (Available online at
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reagent kits on other systems.

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