CAT. NO.

ITP02006
ADVANCED QUALITY

One Step Anti-HIV (1&2) Test

(Whole blood / Serum / Plasma)

FOR IN VITRO DIAGNOSTIC USE

MATERIALS REQUIRED BUT NOT PROVIDED

Positive and negative controls

STORAGE AND STABILITY

INTENDED USE
THE ADVANCED QUALITY ONE STEP ANTI-HIV (1&2) TEST IS A
COLLOIDAL GOLD ENHANCED, RAPID IMMUNOCHROMATOGRAPHIC
ASSAY FOR THE QUALITATIVE DETECTION OF ANTIBODIES TO HUMAN
IMMUNODEFICIENCY VIRUS (HIV) IN HUMAN WHOLE BLOOD, SERUM OR
PLASMA FROM ALL HUMAN GROUPS (INCLUDING NEONATES, PREGNANT
WOMEN, ETC). THIS TEST IS A SCREENING TEST, AND ALL POSITIVES
MUST BE CONFIRMED USING AN ALTERNATE TEST SUCH AS WESTERN
BLOT. THE TEST IS INTENDED FOR HEALTHCARE PROFESSIONAL USE
ONLY.
SUMMARY
The human immunodeficiency virus (HIV) is the causative agent of acquired
1
immune deficiency syndrome (AIDS) . The general method of detecting infection
with HIV is to observe the presence of antibodies to the virus by an EIA method
2
followed by confirmation with Western Blot . The Advanced Quality One Step
Anti-HIV (1&2) Test is a simple, visual qualitative test that detects antibodies in
human whole blood, serum or plasma. The test is based on immunochromatography and can give a result within 15 minutes.
PRINCIPLE OF THE PROCEDURE
The assay starts with a sample applied to the sample well. A recombinant HIV
antigen conjugated to colloidal gold embedded in the sample pad reacts with the
HIV antibody present in whole blood, serum or plasma forming conjugate/HIV
antibody complex. As the mixture is allowed to migrate along the test strip, the
conjugate/HIV antibody complex is captured by recombinant HIV antigen
immobilized on a membrane forming a colored test band in the test region. A
negative sample does not produce a test band due to the absence of colloidal gold
conjugate/HIV antibody complex. The antigens used in the conjugate test are
recombinant proteins that correspond to highly immunoreactive regions of HIV1
and HIV2. A colored control band in the control region appears at the end of test
procedure regardless of test result. This control band is the result of colloidal gold
conjugate binding to the anti-HIV antibody immobilized on the membrane. The
control band indicates that the colloidal gold conjugate is functional.
REAGENTS AND MATERIALS SUPPLIED
Test cards individually foil pouched with a desiccant.
Plastic dropper
Sample diluent
Safety lancet
Alcohol swab
Package insert
1

All the kit reagents can be stored at 2 - 30C in the sealed pouch and under
dry conditions until expiration date.

WARNINGS AND PRECAUTIONS

1. For in vitro diagnostic use only.
2. Treat all samples as though potentially infectious.
3. Wear gloves and protective clothing when handling samples.
4. Operate according to standard safety precautions when dispose
biohazardous materials.
5. Clean and disinfect all spills of samples using a suitable disinfectant, such as
3
1% Sodium Hypochlorite .
6. Sterilize all kit reagents used in this assay prior to disposal.
7. Do not use kit reagents beyond their expiration dates.
8. Do not interchange kit reagents from different lots.
9. Do not re-use the test card or any single use accessories.
Do not use the test card that has become wet or if the foil pouch is damaged.
ASSAY PROCEDURES FOR FINGER BLOOD
1.
2.

3.

Bring the HIV test card, sample diluent, alcohol swab, safety lancet, plastic dropper to
room temperature.
Take out the test card from the sealed pouch.
To perform the HIV test, please follow the steps closely as follow (from picture 1 to
picture 8).

Clean the finger

with alcohol swab

Draw up blood into

the plastic dropper

Bring the safety lancet,

and then twist the knob

Pull out the knob

Add one drop (30l)

of blood

Add one drop (50l)

of sample diluent to
the sample well

Place the activated safety

lancet firmly on the finger.
Push the trigger to prick
the finger

Wait for 15 minutes to

interpret the results

ASSAY PROCEDURE FOR VENOUS BLOOD COLLECTED BY REGULAR

CLINICAL LABORATORY PROCEDURES
SAMPLE COLLECTION
VENIPUNCTURE WHOLE BLOOD
1. Use standard venous phlebotomy procedure to collect a whole blood sample by a tube
containing any of the following anticoagulants: EDTA, heparin, or sodium citrate. Other
anticoagulants have not been tested and may give an incorrect result. If the samples
are not tested at the time of collection, the whole blood can be stored at 2-8 for 3
days. Before testing, mix the blood tube gently by inversion several times to ensure a
homogeneous sample.
2. Pick up an unused sample collection plastic dropper to collect the drop of blood.
3. Using the blood samples in the long-term keeping more than 3 days may cause
non-specific reaction.
SERUM OR PLASMA
1.
SERUM
Use the standard venous phlebotomy procedure to collect a whole blood sample by a
tube NOT containing any of the following anticoagulants: EDTA, heparin, or sodium
citrate. Leave to settle for 30 minutes for blood coagulation and then centrifuge the
blood to get serum sample of supernatant.
2.
PLASMA
Use the standard venous phlebotomy procedure to collect a whole blood sample by a
tube containing any of the following anticoagulants: EDTA, heparin, or sodium citrate.
And then centrifuge the blood to get a plasma sample.
Note:
1.
If serum or plasma samples are not tested immediately, they should be refrigerated at
2-8. For storage period longer than 7days, freezing is recommended. Please bring the
samples to room temperature before testing.
2.
Serum or plasma samples containing a precipitate may yield inconsistent test results.
Such samples must be clarified before testing.

ASSAY PROCEDURE
Do not open pouch until you are ready to test the sample.
1.
2.
3.
4.

5.

Bring all reagents and samples to room temperature.

Remove the test card from the foil pouch and place on a clean dry surface.
Identify the test card for each sample or control.
Dispense one drop (30l) of sample or control into the sample well on the card using
the plastic dropper provided, then add one drop (50l) of sample diluent into the same
well.
Interpret test results at 15 minutes.

Negative

Positive

Invalid

30ul (1 drop) of specimen

50 ul(1 drop) of diluent

Notes:
1. The positive results could appear as soon as 1 minute for a sample with high
levels of HIV antibodies.
2. Do not interpret result after 20 minutes.
INTERPRETATION OF TEST RESULTS
1. Positive: Both purplish test band and purplish control band appear on the membrane.
2. Negative: Only the purplish control band appears on the membrane. The absence of a
test band indicates a negative result.
3. Invalid: There should always be a purplish control band in the control region regardless
of test result. If control band is not seen, the test is considered invalid. Repeat the test
using a new test test.
Note: It is normal to have a slightly lightened control band with very strong positive samples as
long as it is distinctly visible.
PERFORMANCE CHARACTERISTICS
The performance of the Advanced Quality ONE STEP Anti-HIV (1&2) Test has been
evaluated by testing samples from random blood donors, from patients with HIV infection
and commercial seroconversion panels. The performance evaluations were conducted in to
show compliance to the Common Technical Specifications including studies performed
in-house and in two external laboratories in Europe.
1.

Sensitivity
Table I.

Performance on HIV positive samples

One Step anti-HIV test

Types

Total samples

Positive by

Sensitivity

HIV-1 positive with reference tests

259

260

99.62%

HIV-1 positive from EDTA plasma /

whole blood comparison study

100

100

100%

HIV-1 non-B subtype *

40

40

100%

HIV-2

100

100

100%

Total

499

500

99.8%

*Subtypes of A, C, D, F, G, H, J, K, O and CRF AE etc. were included.

Negative

One Step Anti-HIV Test

PRB-914 N

5/5

5/5

PRB-916 P

2/6

2/6

PRB-919 S

2/3

2/3

PRB-924 X

3/8

3/8

PRB-925 Y

2/6

2/6

PRB-926 Z

2/6

2/6

PRB-927 AB

4/5

3/5

PRB-934 AI

2/3

2/3

PRB-942 AR

0/4

0/4

PRB-947 AW

3/4

3/4

PRB-950 AZ

1/4

1/4

PRB-951 BA

1/6

1/6

PRB-952 BB

2/6

2/6

PRB-953 BC

1/4

1/4

PRB-954 BD

0/7

0/7

PRB-955 BE

2/5

1/5

PRB-956 BF

0/5

0/5

PRB-957 BG

1/7

1/7

PRB-958 BH

2/6

2/6

PRB-959 BI

4/7

4/7

Total score

39/107

37/107

The following results were obtained with the One Step Anti-HIV Test and with the
reference method Enzygnost Anti-HIV Plus. Among the 107 samples of the
sero-conversion panels, 31 samples were defined as sero-conversion HIV samples,
and all yielded a positive result with the One Step anti-HIV test; 16 samples were
determined as early seroconversion HIV samples, and only two were missed
compared to the reference.
2.

1000

1000

1000

* Reference test.
Besides, 200 clinical samples, 200 samples from pregnant women (including 180 of
1st pregnancy and 20 of 2nd or later pregnancy) were tested; all the results indicated
a specificity of 100%.
3.

Potential interference
Table IV. Performance on potentially interfering samples
One Step anti-HIV Test

Interfering samples
Negative

positive

Total

anti-HCV positive

20

20

anti-HBsAg positive

20

20

anti-HBc positive

20

20

anti-HTLV I/II positive

20

20

anti-HEV positive

20

20

anti-CMV IgM

10

anti-EBV IgM

10

10

Rheumatoid factor

10

10

Total

129

130

The results showed Specificity on possibly interfering samples of 99.23%.

4.

Performance on deferent types of samples

Sensitivity and specificity tests above were performed on EDTA plasma samples.
Compatibility of EDTA plasma and whole blood samples can be concluded from
above tests, further more, The One Step anti-HIV test can be performed on other
sample types as well: citrate plasma, heparin plasma, serum etc.
Table V. Performance on deferent types of samples
One Step anti-HIV Test
EDTA plasma samples

Specificity

Whole blood samples

Negative

Table III. Performance on negative blood donations

Total

1000

Prism HIV combo

Number of reactive panel members / total number of panel members

Enzygnost anti-HIV Plus

positive

EDTA plasma samples

Prism HIV combo*

Table II. Performance on Sero-conversion panels

Panel ID

One Step anti-HIV Test

blood donor samples

The diagnostic sensitivity of ONE STEP Anti-HIV on HIV positive samples is

calculated to be 99.8%. For the sample lost, further characterization revealed that it
was subtype A virus of recent infection close to sero-conversion. And it is lost also by
other two CE-marked rapid tests.

Citrate plasma samples

positive

Total

negative

30

30

positive

30

30

Heparin plasma
samples

serum samples

total

30

30

60

negative

30

30

positive

30

30

total

30

30

60

negative

30

30

positive

30

30

total

30

30

60

It is concluded that the ONE STEP Anti-HIV test plays the same effect on the all
sample types.
LIMITATIONS
1.
2.
3.
4.
5.
6.

7.
8.

The assay should be performed in normal room temperature (10-30).

Test card should be used immediately after being taken from the package (within 2
hours).
The kit reagents may be stored under 2-30 and dry condition. If refrigerated, the
reagents should be brought to room temperature(10-30) before testing.
Only samples that are not hemolyzed and that are with good fluidity can be used in this
test.
Do not agitate the sample. Insert a pipette just below the surface of the sample to
collect the sample.
Fresh samples are best but refrigerated samples can also be used after balancing to
the room temperature. Frozen serum and plasma can be used after thawing, but frozen
whole blood samples cannot be used in this test.
Although the test is very accurate, a low incidence of false results may occur, so all
positive results must be confirmed by an alternative method.
If questionable results are obtained, the test should be repeated on a fresh whole blood
sample using a new test.

BIBLIOGRAPHY
1.

Blattner, W., Gallo, R.C. and Temin. H.M. HIV causes AIDS. Science. 241:515, 1988.

2.

Curran, J.W., Morgan. W.M., Hardy, A.M., et al. The epidemiology of AIDS: Current status and
future prospects. Science 1985; 229:1352:1352-7.

3.

Centers for Disease Control. Recommendation for prevention of HIV transmission in health care
setting. MMWR 36, Supplement No. 2S, 1987.

InTec PRODUCTS, INC.

39801