70086877 10/3/03 9:48 AM Page 1

M i c ro e l i s a s y s t e m

Vironostika
®

HIV Uni-Form II Ag/Ab

70086877 10/3/03 9:48 AM Page 3

Vironostika HIV Uni-Form II Ag/Ab

®

Table of contents

1 Product details
2 Introduction
3 Principle of the procedure
4 Contents of the kit
5 Additional materials and instrumentation required
6 Type of specimens, handling and storage
Specimen collection
Specimen handling and storage
7 Personal safety
8 Reagent preparation
Microelisa strips
Phosphate buffer
TMB substrate
Sulfuric acid
9 Storage conditions and stability of reagents
10 Procedural precautions
11 Assay procedure
12 Results
Manual calculation
Calculation example
Interpretation of results
13 Automated processing
14 Performance characteristics
Diagnostic sensitivity
Diagnostic specificity
Analytical sensitivity
Analytical specificity
Precision
Interfering factors
Limitations of the assay
15 Availability
16 Explanation of the symbols
17 Schematic representation of assay procedure
References

® Registered trademark in one or more countries

70086877 10/3/03 9:48 AM Page 4

1 Product details
Manufacturer: bioMérieux bv, Boseind 15, 5281RM Boxtel, The Netherlands

Intended use: In vitro diagnostic medical device. Vironostika HIV Uni-Form II Ag/Ab is an
enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies
to human immunodeficiency virus type 1 and/or 2 (anti-HIV-1, anti-HIV-2 and
anti-HIV-1 group O) and HIV-1 antigen in human serum or plasma and is
indicated:
– for the screening of donated blood,
– as an aid in assessing the clinical condition of persons with an unknown
status related to infection with HIV-1 and/or HIV-2.

Use by: Use before the end of the indicated month. See labels for the expiry date of
individual kit components (date = YYYY-MM).

Storage: Store between +2 and + 8 °C.

Special precautions: This kit contains substances of both human and animal origin.
All such materials should be handled with caution and treated as being
potentially infectious.

2 Introduction
The human immunodeficiency viruses type 1 and type 2 are etiological agents of the acquired
immunodeficiency syndrome (AIDS) and related conditions. HIV has been isolated from patients with
AIDS, AIDS related complex (ARC) and from healthy individuals at high risk for AIDS. Infection with
HIV is followed by an acute illness that has flulike characteristics. This phase may remain un-noticed
and the relationship to HIV infection may not be clear in many cases. The acute phase is
typically followed by an asymptomatic carrier state, which progresses to clinical AIDS in about 50 % of
infected individuals within 10 years after seroconversion.(1)

Serological evidence of HIV infection may be obtained by testing for HIV antigens or antibodies in
serum or plasma of individuals suspected of HIV infection. Antigens can generally only be detected
during the acute phase and during the symptomatic phase of AIDS. Antibodies to HIV-1 and/or HIV-2
can be detected throughout virtually the entire infection period, starting at or shortly after the acute
phase and lasting till the end stage of AIDS. The use of highly sensitive antibody assays is therefore an
established approach in serodiagnosis of HIV infection and in the screening of blood and blood prod-
ucts. Progressive improvements in assay sensitivity have reduced the so-called window phase, i.e. the
time between infection with the HIV virus and the moment that antibodies to HIV can be detected by
sensitive HIV antibody tests. Further shortening of this window period can be achieved by the incor-
poration of HIV antigen detection in such HIV antibody tests enabling the detection of infected individ-
uals at the earliest possible moment.

3 Principle of the procedure

Vironostika HIV Uni-Form II Ag/Ab is an ELISA based on a one-step “sandwich” principle.
A mixture of HIV antigens and HIV antibodies coupled to horseradish peroxidase (HRP)
serves as the conjugate with tetramethylbenzidine (TMB) and peroxide as the substrate. Upon com-
pletion of the assay, the development of color indicates the presence of HIV antibody or HIV antigen,
while no or low color development suggests the absence of HIV antibodies or antigens.

Specifically, microelisa wells are coated with HIV-1 gp160 (2), HIV-1 ANT70 peptide (3,4), HIV-2 env pep-
tide (amino acids 592-603) (5) and anti-HIV-1 p24. Each microelisa well contains an HRP-labeled con-
jugate sphere of the same HIV-antibody/antigen mixture. The specimen diluent, which is added to the
wells first, will dissolve the conjugate sphere.
Then the test sample or appropriate control containing HIV antibody or antigen is incubated in the
microelisa wells. If HIV-1 and/or HIV-2 antibody is present in the sample a solid phase antigen/anti-
HIV/enzyme labeled antigen complex is formed. If HIV-1 antigen is present in the sample, a solid
phase antibody/HIV antigen/enzyme labeled antibody complex is formed. Following a wash proce-
dure and incubation with TMB substrate, color develops which turns yellow when the reaction is
stopped with sulfuric acid. If anti-HIV-1, anti-HIV-2, anti-HIV-1 group O and/or HIV antigen is present in
70086877 10/3/03 9:48 AM Page 5

the sample, an intense color develops. However, when the sample is free of anti-HIV and HIV antigen,
no or low color forms with the addition of substrate.

cf g
c j H2SO4
c D h
Solid phase anti-HIV-1/ HRP-labeled TMB
HIV-1/HIV-2/ anti-HIV-2/ mixture of substrate
HIV-1 group O/ anti-HIV-1 group O/ HIV-1/HIV-2/
anti-HIV-1 HIV-1 antigen HIV-1 group O/
mixture in sample anti-HIV-1

Incubate 60 min at 37 °C Incubate Stop and read

30 min at (450 nm)
15 to 30 °C

4 Contents of the kit

192 test kit 576 test kit Components ff
2x 6x Microelisa strip plates n
12 strips per plate, each with 8 wells coated with a mixture of HIV-1 gp160, HIV-1 ANT70,
HIV-2 env (aa 592-603) and anti-HIV-1 p24. Each well contains a pearlshaped sphere,
containing lyophilized HRP-labeled HIV-1 gp160, HIV-1 ANT70 and HIV-2 env (aa 592-603)
conjugate and anti-HIV-1 p24 (murine monoclonal).
Prepare for use as described in section 8. Note: The strips can be broken in the middle to
be used as 4-well-strips. The foil label contains 4 peel off strip plate identification labels in
bar code format. A new label should be stuck on the strip holder for each separate test run.

1x 1x Negative control -4
Human serum nonreactive for anti-HIV and HIV antigen.
Preservatives: 0.1 g/l gentamicin sulfate and 0.2 ml/l cinnamaldehyde.
Ready to use as supplied. Contents: 3.0 ml

1x 1x Anti-HIV-1 positive control A1

Human serum containing human monoclonal anti-HIV-1.
Preservatives: 0.1 g/l gentamicin sulfate and 0.2 ml/l cinnamaldehyde.
Ready to use as supplied. Contents: 2.0 ml

1x 1x Anti-HIV-2 positive control B2

Human serum containing murine monoclonal anti-HIV-2.
Preservatives: 0.1 g/l gentamicin sulfate and 0.2 ml/l cinnamaldehyde.
Ready to use as supplied. Contents: 2.0 ml

1x 1x HIV-1 antigen positive control C3

HIV-1 p24 (inactivated).
Preservatives: 0.1 g/l gentamicin sulfate and 0.2 ml/l cinnamaldehyde.
Ready to use as supplied. Contents: 2.0 ml

1x 3x Specimen diluent H8
Contains stabilizing protein and detergent.
Preservative: 0.2 % Kathon CG.
Ready to use as supplied. Contents: 28 ml

1x 1x Phosphate buffer concentrate O7

Dilute 25-fold in distilled or deionized water as described in section 8. Contents: 100 ml

2x 4x TMB solution I0
Tetramethylbenzidine in citric acid.
Preservative: 1 g/l 2-chloroacetamide. Combine with urea peroxide solution as described
in section 8. Contents: 11 ml

2x 4x Urea peroxide solution LQ

Preservative: 1 g/l 2-chloroacetamide.
Combine with TMB solution as described in section 8. Contents: 11 ml

1+1 1+1 Clamp and rod

Closure for strip plate foil pack.
70086877 10/3/03 9:48 AM Page 6

3x 7x Plate sealers
Perforated, adhesive.

2x 2x Sheets of labels
Protocol, reagent and working solution identification bar code labels for use in auto-
mated processing.

The version code i of this assay is indicated on the label on the box. The microelisa strips, controls and specimen
diluent are assay specific. All components are marked with the assay code, A5, which forms part of the assay version code.

5 Additional materials and instrumentation required

• (Freshly) distilled or deionized water.
• 1 mol/l sulfuric acid (analytical grade).
• Disposable V-shaped troughs.
• Disposable vial(s) (glass or plastic) for preparation of TMB substrate.
• Disposable gloves.
• Timer.
• Sodium hypochlorite solution (5 %) or other suitable disinfectant.
• Appropriate biohazard waste containers for potentially infectious materials.
• Absorbent tissue.
• Dispensing system and/or disposable tip pipettes (single- or multi-channel) capable of dispensing
100 µl ± 5 µl, 50 µl ± 2.5 µl, 1 ml ± 50 µl and 5 ml ± 250 µl and tips.
• Vortex mixer.
• Microshaker for dissolving and mixing conjugate with samples , speed approximately 15 Hz
(= 900 revolutions per minute).
• Incubator capable of heating a microplate and its contents to 37 ± 2 °C within 30 minutes and main-
taining 37 ± 2 °C.
• Microwell aspiration/wash system capable of containing aspirated waste in a closed container and
capable of filling and aspirating the wells completely without overflowing from one well to another.
Preferably the wash system is capable of filling the wells with a fluid volume greater than the well volume
while simultaneously aspirating excess volume in order to avoid overflowing.
• Microwell reader, single wavelength 450 ± 5 nm or dual wavelength 450 ± 5 nm and 620 to 700 nm as
reference with a linear absorbance range of 0 to 2.000 or higher.

Individual instrument manuals provided by the manufacturer should be consulted for additional infor-
mation regarding the following:
– Installation and special requirements.
– Operation principles, instructions, precautions, and hazards.
– Manufacturer’s specifications and performance capabilities.
– Service and maintenance information.
– Quality control procedures.

6 Type of specimens, handling and storage s

Specimen collection
• Human serum or plasma may be used. No special preparation or fasting of the blood donor is neces-
sary. Blood should be collected by normal venipuncture technique and handled with the proper pre-
cautions in accordance with standard laboratory procedures. No adverse effects have been observed
using citrate, heparin or EDTA as anticoagulants. The use of other anticoagulants may affect the out-
come of the assay.
• This assay is not suitable for the testing of pooled specimens.

Specimen handling and storage

• Fresh specimens may be stored for up to one week at 2 to 8 °C if free of microbial contamination.
If longer storage is required, specimens should be stored frozen at –20 °C or below. Conditions that
favor microbial growth should be avoided after thawing. The quality of the specimens can be seriously
affected by microbial growth, which may lead to erroneous results.
• Specimens should not be subjected to more than one freeze/thaw cycle as this can lead to erroneous
results. Heat inactivation at 56 °C for 30 min does not affect the test result.
• Specimens should be equilibrated to room temperature (15 to 30 °C) before testing begins.
• The presence of sodium azide or particulate matter in specimens may lead to erroneous results
• Storage of specimens in self-defrosting freezers is not recommended.
70086877 10/3/03 9:48 AM Page 7

7 Personal safety
• Handle Vironostika HIV Uni-Form II Ag/Ab with care; treat as potentially infectious material.
The viral antigen used for coating the microelisa strips and for the conjugate, and the viral antigen used
for the HIV-1 antigen positive control have been treated to inactivate HIV.
The other components in this kit which are prepared from human serum or plasma have been tested,
and found to be nonreactive for antibody to HIV-1 and HIV-2, antibody to hepatitis C virus (HCV) as well
as for hepatitis B surface antigen (HBsAg). However, as it is not possible to offer complete assurance
that infectious agents are not present, all materials of human origin should be handled as though they
might contain potentially infectious agents.
• Use disposable gloves and handle all materials used in the assay including samples, waste solution,
reaction trays and pipettes, cautiously as though capable of transmitting infectious agents. Consult a
physician immediately in the event that contaminated materials are ingested or come in contact with
open lacerations, lesions, or other breaks in the skin.
• Immediately clean up any spillage containing potentially infectious agents with a 1 : 10 dilution of 5 %
(freshly prepared) sodium hypochlorite. Dispose of the cleaning material by a suitable method.
• Dispose of all specimens and materials used to perform the assay as if they contain infectious agents.
Use one of the following methods to treat waste prior to disposal:
– Autoclave for 60 minutes at 121 °C.
– Incinerate disposable materials.
– Mix liquid waste with 5 % (freshly prepared) sodium hypochlorite solution so that the final concen-
tration is approximately 0.5 % sodium hypochlorite. Allow to stand 30 minutes before disposal.
Caution: Neutralize liquid waste that contains acid before adding sodium hypochlorite.

8 Reagent preparation r
Prepare the following reagents before starting the assay procedure. Reagents and samples should be
equilibrated to room temperature (15 to 30 °C) before beginning the assay and can remain at room
temperature during testing. Store reagents at 2 to 8 °C when not in use. Use clean containers for the
preparation of reagents; after cleaning, rinse container thoroughly with distilled or deionized water
before use.

Microelisa strips n
Bring foil pack to room temperature (15 to 30 °C) before opening to prevent condensation on the
microelisa strips. After opening, the strips are stable for 8 weeks at 2 to 8 °C, provided that the foil
pack is resealed with the clamp and rod provided. Write the expiry date after opening on the label.
The silica gel bag should not be removed from the foil packaging.
The foil label contains four bar code peel-off labels for automated processing of the strip plate.
A new label should be stuck on the strip holder for each separate assay run.

Phosphate buffer MS
Check the phosphate buffer concentrate for the presence of salt crystals. If crystals have formed in the
solution, resolubilize by warming at 37 °C until all crystals have dissolved. Dilute the phosphate buffer
concentrate 1 : 25 with distilled or deionized water, e.g., pour the contents of the buffer concentrate
(100 ml) in a flask and fill up to 2 500 ml with water. Prepare at least 25 ml (plus the priming volume
required by individual instruments) (1 ml concentrated buffer with 24 ml water) of buffer for each
microelisa strip used. Mix well before use. Phosphate buffer is stable for 2 weeks at 2 to 8 °C.
Write the expiry date on the label, which is supplied in the kit.

TMB substrate JP
To prepare the TMB substrate, combine the required amount of TMB solution in a disposable vial in
equal parts with urea peroxide solution according to the number of wells being run (see chart below).
Mix well. Protect TMB solution and TMB substrate from excessive light. TMB substrate must be almost
colorless when used. Note: Solutions containing TMB or urea peroxide should not come into contact
with metal or metal ions since this may give rise to unwanted color formation. TMB substrate is stable
for 8 hours at room temperature (15 to 30 °C) if kept in the dark. Write the time of expiry on the
label, which is supplied in the kit.
70086877 10/3/03 9:48 AM Page 8

I0 + LQ
Number of wells TMB solution Urea peroxide solution
1 – 16 1 ml + 1 ml
17 – 32 2 ml + 2 ml
33 – 48 3 ml + 3 ml
49 – 64 4 ml + 4 ml
65 – 80 5 ml + 5 ml
81 – 96 6 ml + 6 ml

Sulfuric acid G9
Sulfuric acid is corrosive and should be handled with care to prevent exposure to skin and eyes.
Note: When preparing diluted sulfuric acid (1 mol/l) from a concentrated solution, remember that
acid should always be added slowly to water while stirring (e.g., 50 ml concentrated acid (18 mol/l)
to 850 ml distilled or deionized water). A label for the working solution is provided in the kit.

9 Storage conditions and stability of reagents

Store unopened components at 2 to 8 °C.
Components have a limited shelf life after opening and/or preparation:

Microelisa strips 8 weeks 2 to 8 °C in resealed pack

Phosphate buffer 2 weeks 2 to 8 °C after preparation
TMB substrate 8 hours 15 to 30 °C if kept in the dark
All other components 16 weeks 2 to 8 °C in original container

Note: Components, either unopened or opened, may not be used after the expiry date printed on the
label of each component.

10 Procedural precautions
• Check all packaging before using the kit. Damage to the packaging does not prevent the contents of the
kit from being used. However, if the outer packaging is damaged the user must check that components
of the kit are intact before using them.
• Alterations in the physical appearance of assay kit materials may indicate instability or deterioration.
• Do not perform the assay in the presence of reactive vapors (e.g., from sodium hypochlorite, acids,
alkalis, or aldehydes) or dust because the enzymatic activity of the conjugate may be affected.
• Strips of the microelisa plate are removable. Store unused strips as described in section 8. Verify that
the conjugate spheres are on the bottom of the wells before removing the strip sealers. Before testing
begins, the user should inspect the microelisa strip holder and ensure that all strips are secure.
Strip holders should be handled with care to ensure that no strip becomes dislodged during testing.
• Microelisa strips may be used only once.
• All reagents and specimens must be mixed well before use.
• To avoid contamination, do not touch the top or the bottom of the strips, the edge of the wells or the
liquid in the wells or the conjugate sphere with fingers or pipette tips.
• All pipetting steps should be performed with the utmost care and accuracy. Cross-contamination between
reagents and samples may invalidate results. Avoid microbial or any other contamination of reagents.
• Remove any bubbles in the well; e.g. by gentle tapping.
• Do not allow the microelisa wells to dry once the assay has begun.
• Prevent evaporation during sample incubation (e.g., by covering the strips with plate sealer. Remove
the plate sealer before commencing washing).
• Routine maintenance of the aspiration/wash system is strongly recommended to prevent carryover
from highly reactive specimens to nonreactive specimens.

11 Assay procedure
1 Fit the strip holder with the required number of microelisa strips. n
Remove the strip sealers. Note: The microwells can be visually inspected to check for the addition of
specimen diluent, samples and controls. Upon addition of specimen diluent the purple conjugate
sphere will dissolve to give a green solution. This solution becomes blue/purple after addition of either
sample or control.
70086877 10/3/03 9:48 AM Page 9

2 Pipette 100 µl specimen diluent into all wells, i.e. including control wells. p
3 Pipette 50 µl sample or control into assigned wells. Include three negative controls and one p
anti-HIV-1 positive control in each strip holder. If desired, one anti-HIV-2 positive control (50 µl)
and/or one HIV-1 antigen positive control may be included in each strip holder. Always pipette the
controls after the samples. If all wells in a strip are not used for sample or control, fill the unused wells
with specimen diluent in order to dissolve the conjugate sphere and prevent malfunction of the
aspiration/wash system.

4 Mix well (e.g., using a microshaker, speed approximately 15 Hz a

(= 900 revolutions per minute) for 15 seconds, or equivalent).

Incubate the strips at 37 °C for 60 ± 5 minutes. tk

5 Wash and soak each well six times with phosphate buffer. d
– Incomplete washing will adversely affect the assay outcome.
– Aspirate the well contents completely into a waste flask. Then fill the wells completely with phosphate
buffer avoiding overflow from one well to another and allow to soak for 30 to 60 seconds.
Aspirate completely and repeat the wash and soak procedure five times for a total of six washes.
– It is preferable to wash the wells by filling them with a fluid volume greater than the well volume
while simultaneously aspirating excess volume in order to prevent overflowing.
– Remove any remaining fluid on the top and the bottom of the microelisa strips and strip holder
after the last aspiration; e.g., by blotting with absorbent tissue.

6 Pipette 100 µl TMB substrate into each well. Do not mix or shake. p
7 Incubate the strips at 15 to 30 °C for 30 ± 2 minutes. tk
8 Stop the reaction by adding 100 µl 1 mol/l sulfuric acid to each well; use the same pipetting h
sequence and time intervals used for TMB substrate addition. Ensure thorough mixing; e.g., by
tapping the side of the strip holder. Plates should be read within 15 minutes.

9 Blank the reader on air, i.e. without strip holder and strips, and read the absorbance of b
the solution in each well at 450 nm (single wavelength) or 450 nm and 620 to 700 nm as
reference (dual wavelength).

12 Results
Manual calculation
Calculations must be made separately for each strip holder.
NC = Absorbance of the negative control
PC1 = Absorbance of the anti-HIV-1 positive control
PC2 = Absorbance of the anti-HIV-2 positive control
PC3 = Absorbance of the HIV-1 antigen positive control

Qualification of NC values
1 NC must be < 0.250. Eliminate any NC ≥ 0.250.
2 Determine the mean (NCx) value of the remaining controls.
3 NC must be ≤ 1.4NCx. Eliminate any NC > 1.4NCx and recalculate NCx.
4 NC must be ≥ 0.6NCx. Eliminate any NC < 0.6NCx and recalculate NCx.
5 Repeat steps 3 and 4 until no more outliers are found.

Assay validity
An assay run is valid if
1 more than half the number of the negative controls remain;
2 PC1 – NCx ≥ 0.600;
3 PC2 – NCx ≥ 0.600 (if used);
4 PC3 – NCx ≥ 0.400 (if used).

Cut-off value
If the test run is valid, calculate the cut-off value NCx + 0.100.

A test sample is reactive if sample absorbance is ≥ cut-off value.

70086877 10/3/03 9:48 AM Page 10

10

A test sample is nonreactive if sample absorbance is < cut-off value.

Calculation example
NC = 0.089; 0.096; 0.088 NCx = 0.091
PC1 = 1.549
PC2 = 1.523
PC3 = 1.398

Eliminate any aberrant control:

NC ≥ 0.250 None eliminated
NC > 1.4NCx 1.4(0.091) = 0.127 None eliminated
NC < 0.6NCx 0.6(0.091) = 0.055 None eliminated

Ensure the following is within specified acceptance criteria:

PC1 – NCx ≥ 0.600 1.549 – 0.091 = 1.458 Pass
PC2 – NCx ≥ 0.600 1.523 – 0.091 = 1.432 Pass
PC3 – NCx ≥ 0.400 1.398 – 0.091 = 1.307 Pass

Calculate cut-off value:

Cut-off = NCx + 0.100 = 0.091 + 0.100 = 0.191

Interpretation of results
• A nonreactive result indicates that the sample tested either does not contain anti-HIV-1, anti-HIV-2,
anti-HIV-1 group O or HIV-1 antigen or that the sample contains one or more of these below the
detection limit of the Vironostika HIV Uni-Form II Ag/Ab assay.
• A reactive result indicates that the sample tested either contains anti-HIV-1, anti-HIV-2, anti-HIV-1
group O and/or HIV-1 antigen or that it contains a nonspecifically reacting factor.
• Specimens that show an initially reactive result should be retested in duplicate.
Only specimens reactive in one or both retests should be considered reactive for anti-HIV-1, anti-HIV-2,
anti-HIV-1 group O and/or HIV-1 antigen.
Because all highly sensitive immunoassay systems have a potential for nonspecific reactions, repeatedly
reactive specimen results must be verified using an appropriate test method.
Due to the high sensitivity of Vironostika HIV Uni-Form II Ag/Ab in early seroconversion samples, it is
recommended to include a sensitive HIV antigen assay in confirmatory testing.
• Initially reactive specimens that are nonreactive in duplicate testing, should be considered nonreactive.
Nonrepeatable reactive results may be due to one or more of the following technical problems:
– Carry-over from a highly reactive sample due to contamination of equipment or pipette tips.
– Substrate contamination with metal ions.
– Cross-contamination from moisture or reagent droplets.
– Inadequate wash or aspiration during wash procedure.
– Reading errors; e.g. due to liquid droplets under the well or air bubbles in the well.

13 Automated processing
Instruments and software packages, as well as their respective processing and calculation protocols,
are available from bioMérieux for the automated processing and test result calculation of the
Vironostika HIV Uni-Form ll Ag/Ab assay. The protocols that may be used and a summary of the
processing steps for the Vironostika HIV Uni-Form ll Ag/Ab assay are given in the following table.

c t k b
protocol incubate at incubate for read at
A5V50P01 37 °C 60 min 450 nm
A5V50P02 37 °C 60 min 450; 620 - 700 nm
Note: Protocol, reagent and working solution identification bar code labels for use in automated
processing are provided.
70086877 10/3/03 9:48 AM Page 11

11

14 Performance characteristics
Correct performance of this in vitro diagnostic product can only be guaranteed by bioMérieux when it
is used as intended, and conform the instructions for use, and where appropriate in combination with
other in vitro diagnostic medical devices as well as accessories released and/or qualified by bioMérieux.
For additional information, such as guidance for other conditions of use, please contact your local
bioMérieux representative.

The results presented below are those from a representative batch of Vironostika HIV Uni-Form II Ag/Ab
that has been evaluated by bioMérieux Development Laboratories.
These results were obtained using the single wavelength assay procedure.

Diagnostic sensitivity

Table 1: Diagnostic sensitivity in seroconversion panels.

BBI panel first positive sample number

Vironostika 3rd generation
HIV UF-II Ag/Ab HIV antibody test

panel J 2 3
panel P 4 5
panel W 9 9
panel X 5 6
panel Z 4 5
panel AF 6 6
panel AG 4 4
panel AJ 6 7
panel AK 4 6
panel AN 3 5
panel AQ 4 4
panel BA 4 6
panel BE 3 4
panel BG 6 7
panel BH 5 5
panel BI 2 3

Table 2: Diagnostic sensitivity in anti-HIV reactive samples

No. of samples sensitivity

445 anti-HIV-1 100 %
200 anti-HIV-2 100 %
303 HIV antigen 100 %

Diagnostic specificity

Table 3: Diagnostic specificity in random donor samples (Serum and EDTA-plasma)

No. of donor samples specificity

4796 99.9 %

Analytical sensitivity
Not applicable.

Analytical specificity
The following samples were all found to be negative in the assay:
70086877 10/3/03 9:48 AM Page 12

12

Table 4: Analytical specificity

Samples containing Number negative positive

tested result result
Anti-nuclear antibodies 20 20 0
Rheumatoid factor 20 20 0
HAMA 25 25 0
Epstein-Barr virus IgM 10 10 0
Cytomegalovirus IgM 10 10 0
Elevated levels of bilirubin 10 10 0
Elevated levels of lipid 10 10 0
Elevated levels of hemoglobin 10 10 0
Elevated levels of protein 10 10 0
Clinical samples
(various clinical conditions) 400 400 0

Samples known to contain extremely high concentrations of anti-HIV do not interfere with detection;
i.e. there is no relevant high dose hook effect.

Precision

Table 5: Precision

Variation Control type CV %

Reproducibility negative control 14.7 %
anti-HIV-1 positive control 14.7 %

Interfering factors
The presence of sodium azide or particulate matter in specimens may lead to erroneous results.

Limitations of the assay

Not applicable

15 Availability
Vironostika HIV Uni-Form II Ag/Ab

Number of tests per kit Catalogue number

a r
192 285046 C 0543
576 285047

For technical assistance, contact your local bioMérieux representative.

70086877 10/3/03 9:48 AM Page 13

13

16 Explanation of symbols

- Reagent symbol for Negative control

A Reagent symbol for Anti-HIV-1 Positive control
B Reagent symbol for Anti-HIV-2 Positive control
C Reagent symbol for HIV-1 Antigen Positive control
H Reagent symbol for Specimen Diluent
O Reagent symbol for 25x Phosphate buffer concentrate
M Reagent symbol for Phosphate buffer
I Reagent symbol for TMB solution
J Reagent symbol for TMB + UP substrate
L Reagent symbol for Urea peroxide solution
G Reagent symbol for Sulfuric acid 1 mol/l
4 Reagent code for Negative control
1 Reagent code for Anti-HIV-1 Positive control
2 Reagent code for Anti-HIV-2 Positive control
3 Reagent code for HIV-1 Antigen Positive control
8 Reagent code for Specimen Diluent
7 Reagent code for Phosphate buffer concentrate
S Reagent code for Phosphate buffer
0 Reagent code for TMB solution
Q Reagent code for Urea peroxide solution
P Reagent code for TMB/UP substrate
9 Reagent code for Sulfuric acid (1 mol/l)
i Assay version
ff Unopened Reagents
n Microelisa strip plate
s Specimen
r Reagent preparation/working solution
d Wash procedure
c Processing and calculation protocol
b Read at … nm
p Pipette … µl
a Shake for … sec
k Incubate for … min
t Incubate at ….°C
h Stop the reaction
l Batch code
I In vitro diagnostic medical device
t Storage temperature limitations
x Use by
a Contains sufficient materials for <n> tests
C Mark of CE conformity
i Consult instruction for use
M Manufacturer
F Symbol for specific German recycling firm
r Catalogue number

17 Schematic representation of assay procedure

See cover.
70086877 10/3/03 9:48 AM Page 127

1 2 3
Foil pack closure. Fold open end of foil pack over rod. Apply clamp.
Fermeture du sachet. Replier l’extrémité ouverte de l’emballage sur la tige. Appliquer la pince.
Verschließen der Folienpackung. Das offene Ende der Folienpackung über den Stab falten und die Klammer darüber-
ziehen.
Cierre del envase. Doblar el extremo abierto del envase sobre la varilla. Aplicar la pinza.
Chiusura per l’involucro dei supporti. Ripiegare l’estremità aperta dell’involucro sul bastoncino. Applicare il morsetto.
Fechamento da embalagem. Dobre a extremidade aberta sobre o bastão. Em seguida coloque a presilha.
Sluiting van de folieverpakking. Vouw de open kant van de folieverpakking om het staafje. Breng de klem aan.
Lukning af foliepakningen. Buk foliepakningens åbne ende omkring stangen og skub klemmen på efterfølgende.
Förslutning av folieförpackning. Vik den öppna ändan av folieförpackningen över staven. Dra över klämman.
Zamknięcie foliowego woreczka. Zagiąć otwarty koniec foliowego woreczka na pręcie. Nałożyć zacisk.
Ξανασφρ)γισµα της αλουµιννιας συσκευασας προσταας. ∆ιπλ2στε το ανοιχτ κρο της αλουµιννιας
συσκευασας επνω απ τη ρβδο. Eφαρµτε το σωλνα σφργιοης.
70086877 10/3/03 9:48 AM Page 128

Vironostika ®

iHIV Uni-Form II Ag/Ab

8 n 2...8 °C  15...30 °C r
1 O7 + 24 H2O MS
=

1 I0 + 1 LQ = JP

17 H2O + 1 H2SO4 (18 mol/l) = G9

11 1 m n = ..

2 H8 100 µl p
3 s 50 µl p
-4 3 x 50 µl
A1 1 x 50 µl

B2 1 x 50 µl
C3 1 x 50 µl

4 15 s 37 °C 60 ± 5 min atk
5 MS 6x d
6 JP 100 µl p
7 15...30 °C 30 ± 2 min t k
8 G9 100 µl h

 
9 450 nm 450; 620 - 700 nm b 70086877
2003-04

bioMérieux bv
Boseind 15, 5281RM Boxtel, The Netherlands
The logo is a registered trademark and the exclusive property of bioMérieux or one of its affiiliates