HIV303

30 Tests per Kit

HIV 1/2 STAT-PAK® DIPSTICK

A Qualitative Screening Test Kit for the Detection of Antibodies to HIV 1/2 in Human Serum, Plasma or Whole Blood

FOR EXPORT ONLY

FOR IN VITRO DIAGNOSTIC USE
FOR PROFESSIONAL USE ONLY
READ INSTRUCTIONS FOR USE CAREFULLY BEFORE PERFORMING TEST
INTENDED USE PRINCIPLE OF TEST
The Chembio HIV 1/2 STAT-PAK DIPSTICK assay is a single The Chembio HIV1/2 STAT-PAK DIPSTICK assay employs a
use, immunochromatographic screening test which uses a cocktail combination of antibody binding protein, which is conjugated to
of antigens to detect antibodies to HIV 1 and 2 in serum, plasma colloidal gold dye particles, and antigens to HIV1/2, which are
or whole blood. Reactive results are supportive evidence of bound to the membrane solid phase. The sample being tested and
exposure to HIV 1/2 and can be used to support a clinical Running Buffer are applied to the sample pad. The Running
diagnosis of HIV 1 or HIV 2. Nonreactive results, however, Buffer facilitates the lateral flow of the specimen through the
should not be used to exclude infection with HIV 1 or 2. membrane and promotes the binding of antibodies to the antigens.
If present, the antibodies bind to the gold conjugated antibody
SUMMARY AND EXPLANATION
binding protein. In a reactive sample, the dye conjugated-immune
Discovered in 1983, Human Immunodeficiency Virus (HIV) is a complex migrates on the nitrocellulose membrane and is captured
retrovirus identified as the etiologic agent for Acquired by the antigens immobilized in the TEST area producing a
Immunodeficiency Syndrome (AIDS) [1]. AIDS is characterized pink/purple line. In the absence of HIV 1/2 antibodies, there is no
by changes in the population of T-cell lymphocytes that play a key pink/purple line in the TEST area. The sample continues to
role in the immune defense system. In the infected individual the migrate along the membrane and produces a pink/purple line in
virus causes a depletion of subpopulation of T-cells, called T-
the CONTROL area containing immunoglobulin G antigens. This
helper cells, which leaves these patients susceptible to
procedural control serves to demonstrate that specimens and
opportunistic infections and certain malignancies. The major
reagents have been applied properly and have migrated through
routes of transmission are sexual contact, contamination by blood
or blood products and mother-to-newborn transmission [2-4]. the device.

Although there have been decreases in the rate of infection in MATERIALS AVAILABLE AS AN ACCESSORY TO
certain countries, the number of persons infected with HIV THE KIT
globally has continued to increase. By the end of 2005 there were Chembio Rapid HIV 1/2 Antibody Test Kit Controls (Catalogue
approximately 40.3 million people living with HIV/AIDS, up from
HIV104). Each package contains:
about 37.5 million in 2003. Nearly 5 million people were newly
• HIV 1 Reactive Control
infected with HIV/AIDS in 2005. In the same year more than 3
million died of AIDS-related illnesses; more than 500,000 of these • HIV 2 Reactive Control
were children [5]. • Non Reactive Control
• Product Insert for the HIV104 Kit Controls
The HIV virus consists of a genomic RNA molecule protected by
a capsid and an envelope. The HIV envelope is the major target MATERIALS PROVIDED
for humoral antibody response. The presence of the virus in Each kit contains the items to perform 30 tests:
patients causes the immune system to elicit the production of anti- • 30 HIV 1/ 2 STAT-PAK DIPSTICK test strips
HIV antibodies. The detection of these antibodies can be used as a • 1 Running Buffer bottle (8 ml)
diagnostic tool.
• 30 Disposable Sample Loops (5 !L)
ELISA, Western Blots, PCR-based assays and various other test • 30 Backing Cards
systems are currently available for HIV1/2 detection [6-10]. The • 1 Product Insert
Chembio HIV 1/2 STAT-PAK DIPSTICK assay is a rapid MATERIALS REQUIRED BUT NOT PROVIDED
immunochromatographic test, which is simple and easy to use.
• Clock, watch or other timing device
The Chembio HIV 1/2 STAT-PAK DIPSTICK assay utilizes
• Automatic pipettor capable of delivering a 5 !L sample may
immobilized antigens for the detection of antibodies to HIV 1 and
2 in serum, plasma or whole blood with high degree of sensitivity be used in lieu of the disposable 5 !L sample loop supplied
and specificity. with the kit, for other than fingerstick specimens
• Disposable gloves
• Sterile Gauze (for fingerstick whole blood specimens)
• Sterile Alcohol swab (for fingerstick samples only)
• Biohazard disposal container
• Collection devices for samples other than fingerstick
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WARNINGS 4. Always check expiration date prior to testing.
For in-vitro diagnostic use 5. Do not use the test beyond the expiration date printed on the
1. Read the Product Insert completely before using this assay. tube containing the test strips.
Follow the instructions carefully as not doing so may result in 6. Strips should be used immediately following removal from
inaccurate test results. the storage tube.
2. Use of the test kit with specimen types other than those 7. Do not mix reagents from different lot numbers of kits.
specifically approved for use with this device may result in 8. Adequate lighting is required to read the test results.
inaccurate test results.
3. This test should be performed at 18-30°C (64-86°F). If stored SPECIMEN COLLECTION
refrigerated, ensure that the test device is brought to
operating temperature before performing testing. The Chembio HIV 1/2 STAT-PAK DIPSTICK assay can be
4. If the test kit is stored at temperatures outside the storage performed on whole blood, serum or plasma.
temperature 8-30°C (46-86°F), or used outside the operating
Fingerstick Whole Blood:
temperature 18-30°C (64-86°F), use the Kit Controls
Prepare to perform the fingerstick blood collection procedure.
(Catalogue HIV104) to ensure proper performance of the test.
Clean the finger of the person being tested with an antiseptic wipe.
5. Individual(s) infected with HIV-1 and/or HIV-2 who is
Allow the finger to dry thoroughly or wipe dry with a sterile gauze
receiving highly active antiretroviral therapy (HAART) may
pad.
produce negative results.
6. Sample loops are single use only. Discard sample loops
Using a sterile lancet, following laboratory procedure, prick the
following initial use.
finger just off the center of the finger and wipe away the first drop
with sterile gauze. Avoid squeezing the fingertip to accelerate
STORAGE AND STABILITY bleeding as this may dilute the blood with excess tissue fluid.
The HIV1/2 STAT-PAK DIPSTICK test strips should be stored at
temperatures between 8-30ºC in the original vial with desiccant. Collect the sample from the second drop by touching the
The Running Buffer should be stored at 8-30ºC. disposable sample loop provided to the drop of blood until the
sample loop is full. Test immediately, following test procedure
The vial containing the test strips should only be opened to instructions.
remove dipsticks for testing and should be closed immediately and
tightly thereafter. Venous Whole Blood:
Draw blood following laboratory procedure for obtaining venous
NOTE: Do not use test kits beyond the expiration date imprinted blood. Depending on use, collect sample in a tube containing
on the box label and/or vial. citrate, heparin or EDTA. Be sure the tube of blood is well mixed
before sampling. Dip the sample loop into the blood and allow it
CAUTION: DO NOT FREEZE TEST KITS to fill. Follow test procedure instructions.

PRECAUTIONS If tested the same day, venous whole blood may be kept at room
Safety Precautions temperature. Venous whole blood may be stored for up to 3 days
1. Do not eat, drink or smoke in the area where specimens and between 2-8°C (36-46°F) before testing.
kit reagents are handled. Avoid any contact between hands,
eyes or mouth during specimen collection and testing. DO NOT FREEZE WHOLE BLOOD! Allow for refrigerated
2. Wear protective clothing such as laboratory coats, disposable sample to reach room temperature and mix gently before testing.
gloves and eye protection when handling patient specimens.
3. Dispose of all specimens and materials used in the test Serum or Plasma:
procedure in a biohazard waste container. Lancets should be Draw blood following laboratory procedures for obtaining serum
placed in a puncture-resistant container prior to disposal. The or plasma specimens. Collect specimen in a clean tube not
recommended method of disposal of biohazard waste is containing any anticoagulant (serum) or in a tube containing
autoclaving for a minimum of 1 hour at 121ºC. Disposable citrate, heparin or EDTA (plasma), following standard laboratory
materials may be incinerated. Liquid wastes may be mixed procedures for handling and collecting specimens. All specimens
with appropriate chemical disinfectants. A freshly prepared should be centrifuged and handled following local clinical or
solution of 10% bleach (0.5% solution of sodium laboratory procedures. Remove the serum from the clot as soon as
hypochlorite) is recommended. Allow 60 minutes for possible to avoid hemolysis.
effective decontamination.
NOTE: Do not autoclave solutions that contain bleach. Patient samples perform best when tested immediately after
4. For additional information refer to: World Health collection. If not to be tested immediately, specimens should be
Organization (WHO) Rapid HIV Tests: Guidelines for use in refrigerated following collection at 2-8°C and used within 3 days
HIV testing and counseling services in resource-constrained of collection. If testing within 3 days is not possible, the
Settings (2004) [11]. specimens should be frozen (-20ºC or colder).

Handling Precautions NOTE: If specimens are to be shipped, they should be packed in

1. Do not use any test strips if the tube has been deformed. compliance with regulations covering the transportation of
2. Each device is for single use only. etiologic agents.
3. Each sample loop is single use only and must be discarded
following initial use.

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CONFIRMATION OF TEST PERFORMANCE 4. Place the test strip onto the backing card in the marked space
with the green tape facing up and the arrows on the tape
When the test is complete, you will observe a pink/purple line in facing in the same direction as the arrows on the backing
the CONTROL area of the test strip on nonreactive as well as card. Place the backing card and dipstick assembly on a
reactive samples. This serves as an internal control and gives clean, flat surface.
confirmation of proper test performance. Pink/purple lines in both
the TEST and CONTROL areas indicate reactive samples.
4.
TEST PROCEDURE
If specimen to be tested is refrigerated, remove it from the
refrigerator and allow it to come to room temperature prior to
testing.

1. Remove the backing cards and tear off one segment for each
test to be run. Enter patient or specimen ID in the space
marked ID on the card.
1.

5. SPECIMEN COLLECTION
A. Fingerstick Whole Blood
i. Following laboratory procedure, prick
the finger with a lancet and wipe away 5A.
the first drop of blood.
ii. Collect the sample from the second drop
by touching the 5!L sample loop provided
to the drop of blood. (Proceed to Step 6)

B. Serum, Plasma, Venous Whole Blood 5B.

Touch the 5!L sample loop provided to the
sample to fill the circular opening of
the loop with sample.

2. Remove the HIV 1/2 STAT-PAK DIPSTICK test strip from

the vial. The vial with the test strips should only be opened to 6. Holding the sample loop vertically, touch it to the sample pad
remove dipsticks for testing and should be closed tightly in the center of the SAMPLE (S) area of the dipstick to
immediately thereafter. dispense ~5 !L of sample (serum, plasma or whole blood)
onto the sample pad.
3. Being careful not to touch the membrane, turn the test strip so
the green tape is facing away and remove the red liner from 6.
the adhesive strip(s) on the back of the test strip. Discard the
red liner.

3.

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7. Invert the Running Buffer bottle and hold it vertically (not at INTERPRETATION OF RESULTS
an angle) over the sample area. Add 3 drops (~ 105 !L) of
buffer slowly, dropwise, onto the SAMPLE (S) area being Reactive Result
sure each drop is absorbed by the sample pad before adding Two pink/purple lines, one in the TEST area and
the next. one in the CONTROL area indicate a reactive
NOTE: In the case that liquid does not start to flow onto the result. The line in TEST area may look different
membrane within 2 minutes, add one additional drop of from the line in the CONTROL area. The
Running Buffer. intensity of the line in the TEST area will vary
7. with the concentration of specific antibodies,
from barely visible to very dark.

Control Line NOTE: Even a very faint line in the TEST area
Test Line should be considered reactive. It is
recommended that a reactive result be confirmed
by Western Blot or IFA in accordance with the
World Health Organization (WHO)
recommendations. This test is suitable for use in
8. Read the test result between 15 and 20 minutes multi-test algorithms designed for the statistical
after the addition of the Running Buffer. In validation of rapid HIV test results. When
some cases a test line may appear in less than multiple rapid HIV tests are available, this test
should be used in appropriate multi-test
15 minutes however, 15 minutes are needed to
algorithms.
report a nonreactive result. Read results in a
well-lit area. Nonreactive Result
Do not read results after 20 minutes.
One pink/purple colored line in the
CONTROL area, with no colored line in the
NOTE: Discard the used sample loop, test strip, backing card and TEST area indicates a nonreactive result. A
gloves into a biohazard waste container. nonreactive result after 15 minutes indicates
that there are no HIV antibodies in the
patient sample, but this result does not
QUALITY CONTROL exclude HIV infection.
A pink/purple line should always appear in the CONTROL area if Control Line
the test has been performed correctly and the device is working Test Line
properly. It serves as an internal procedural control. *Red Cell Line occurs on rare occasions
with blood specimens, but has no impact on
Good Laboratory Practice (GLP) recommends the use of control
test results. Such a line should not be
materials along with the test samples to ensure proper
interpreted as a positive test result
performance of the test kit. Reactive and nonreactive serum or
plasma based commercial controls should be used for this purpose.
Use controls, as per the TEST PROCEDURE instructions of this
insert. Please note that certain commercial controls designed for Invalid
ELISA may not perform properly with the Chembio HIV 1/2 A pink/purple line should always appear in the CONTROL area,
STAT-PAK DIPSTICK assay. Serum or plasma based controls for no matter if the TEST LINE appears or not. If there is no distinct
example from BBI (Seracare/Boston Biomedica Inc, USA) or pink/purple line visible in the CONTROL area, the test is invalid.
NABI (Nabi Diagnostics, USA) can be used. It is recommended that the test be repeated with a new device.

RUN CONTROLS UNDER THE FOLLOWING

CIRCUMSTANCES:
• Each new operator prior to performing tests on patient
specimens,
• When opening a new kit lot,
• Whenever a new shipment of Kits is received,
• If the temperature of the test storage area falls outside of 8-
30°C (46-86°F),
• If the temperature of the testing area falls outside of 18-30°C Control Line
(64-86°F), Test Line
• At periodic intervals as indicated by the user facility.

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LIMITATIONS OF THE PROCEDURE PRECISION
1. The HIV 1/2 STAT-PAK DIPSTICK procedure and the Intraassay
interpretation of the results must be followed closely. It is a Within run precision was determined by using 10 replicates of two
screening test designed for detecting antibodies against HIV specimens containing different levels of HIV 1/2 antibodies. The
1 and 2 in human serum, plasma or whole blood. Results non reactive and reactive results were correctly identified 100% of
from testing other body fluids or of pooled serum or plasma the time.
should not be used.
Interassay
2. Venous whole blood must be collected with an anticoagulant
Between run precision was determined by using the same two
depending on use. For serum samples collect blood without
specimens in 10 different replicates from three different lots of test
anticoagulant.
devices. Nonreactive and reactive results were observed 100% of
3. Do not open the tube containing test strips until just prior to
the time.
use. Close it immediately after removing required number of
test strips.
Cross Reactivity and Interference
4. Do not use kit contents beyond their labeled expiration date.
No cross reactivity was observed from Hepatitis B, Rheumatoid
5. Ensure finger is completely dry before performing
fingerstick. Factor (~80 IU/ml) and hCG (500 mIU/ml) with the HIV 1/2
STAT-PAK DIPSTICK assay. In addition, no interference from
6. Read results in a well lit area.
bilirubin, hemoglobin or triglycerides was observed.
7. Reactive results must be confirmed with Western Blot or IFA
according to WHO recommendations. The clinical evaluation
of the patient’s situation should be performed before a final REFERENCES
diagnosis is made. A reactive result obtained with a single 1. Essex, M. (1999) Human immunodeficiency viruses in the
rapid test should not be used to diagnose HIV/AIDS infection developing world. Adv Virus Res 53 : 71-88.
even if antibodies to HIV 1 and 2 are present. A nonreactive 2. Kanki, P.J., Hopper, J.R. and Essex, M. (1987) The origins of HIV-1
result at any time does not preclude the possibility of and HTLV-4/HIV/2. Ann N Y Acad Sci 511 : 370-375.
infection with HIV 1/2. 3. Nicoll, A., Gill, O.N. (1999) The global impact of HIV infection and
disease. Commun Dis Publ Health 2 : 85-95.
8. An individual infected with HIV-1 and/or HIV-2 who is 4. Valdiserri R.O., Holtgrave, D.R., West, G.R. (1999) Promoting early
receiving highly active antiretroviral therapy (HAART) may diagnosis and entry into care. AIDS 13 : 2317-2330.
produce false negative results. 5. UNAIDS/WHO Press Release (2005) HIV Infection Rates
9. This assay has not been evaluated for newborn screening, Decreasing in Several Countries but Global Number of People
cord blood specimens, or for individuals less than 13 years of Living with HIV Continues to Rise. Available at
age. www.unaids.org/epi/2005/doc/docs/PR_EpiUpdate_Nov05en.pdf
: 1-3.
6. Essex, M., Kanki, P. J., Marlink, R., et al. (1990) Antigenic
EXPECTED RESULTS characterization of the human immunodeficiency viruses. J Am Acad
This is a qualitative test for the detection of antibodies to HIV 1 Dermatol 22 : 1206-1210.
7. Essex, M., McLane, M.F., Lee, T.H. et al. (1983) Antibodies to cell
and 2 in whole blood, serum or plasma. As described in the
membrane antigens associated with human T-cell leukemia virus in
PERFORMANCE CHARACTERISTICS section below, the patients with AIDS. Science 220 : 859-862.
sensitivity of the Chembio HIV 1/2 STAT-PAK DIPSTICK assay 8. Gallo, R.C., Saluahuddin, S.Z., Popovic, M., et al. (1984) Frequent
was found to be substantially equivalent to EIA and Western Blot detection and isolation of cytopathic retroviruses (HTLV-III) from
tests when tested on selected performance panels. patients with AIDS and at risk for AIDS. Science 224 : 500-503.
9. Kenealy, W., Reed, D., Cybulsky, R., et.al. (1987) Analysis of
PERFORMANCE CHARACTERISTICS human serum antibodies to human immunodeficiency virus (HIV)
using recombinant ENV and GAG antigens. AIDS Res Human
In-house studies demonstrate that the sensitivity of the Chembio Retrovir 3 : 95-105.
HIV 1/2 STAT-PAK DIPSTICK assay is substantially equivalent 10. Kovacs, A., Xu, J., Rasheed, S., et al. (1995) Comparison of a rapid
to EIA and Western Blot tests when tested on BBI performance non-isotopic polymerase chain reaction assay with four commonly
used methods for the early diagnosis of human Immunodeficiency
panels PRZ204, Anti-HIV 1/2 Combo Performance Panel, and
Virus type 1 infection in neonates and children. Pediatr Infect Dis J
PRB204 Mixed Titer Performance Panel and WWRB302 and 14 : 948-954.
WWRB303 Worldwide Performance Panels. 11. World Health Organization (WHO): Rapid HIV Tests: Guidelines for
Use in HIV Testing and Counseling Services in Resource –
Performance data were also obtained in three external studies Constrained Settings. (2004).
using the alternate test procedure in a tube. In a limited evaluation
of 770 samples by the World Health Organization (WHO), the
final sensitivity was 99.0% and the final specificity was 100%
compared to the reference assays. In this study, none of the results
were recorded as indeterminate. Evaluations in Zambia with 236
specimens, (125 reactive / 111 non-reactive) and in Uganda with
503 specimens (90 reactive / 413 non-reactive) both resulted in
100% sensitivity and 100% specificity for the Chembio HIV 1/2
STAT-PAK DIPSTICK assay.

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 SYMBOL LEGEND

CONSULT THE MANUAL BEFORE USE

CAUTION, CONSULT ACCOMPANYING

DOCUMENTS.
DO NOT REUSE

FOR USE WITHIN TEMPERATURE LIMITS

IN VITRO DIAGNOSTIC MEDICAL DEVICE

BATCH CODE

PRODUCT CATALOG NUMBER

MANUFACTURERS IDENTIFICATION

DATE OF MANUFACTURE

USE BY DATE

FOR MORE INFORMATION, CONTACT

M CHEMBIO DIAGNOSTIC SYSTEMS, INC.

3661 HORSEBLOCK ROAD
MEDFORD, NY 11763 USA
Tel : (631) 924-1135
Fax : (631) 924-6033

Email : [email protected]
Web Site : www.chembio.com

Available Accessories
HIV104: HIV Reactive and Nonreactive Controls

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