Performance Characteristics

The 3rd Generation of ONE STEP antibodies to HIV-1/HIV-2 Test 1. Diagnostic sensitivity
HIV type No. of samples tested SD HIV 1/2 3.0
Negative Positive
HIV-1 (*1,2) 480 0 480
HIV-2 (*1,2) 120 0 120
Total 600 0 600 (100%)

2. Diagnostic Sensitivity on Seroconversion panels – seroconversion HIV samples and early HIV seroconversion samples (*7)
[Seroconversion panel sensitivity]
Comparative analysis between SD BIOLINE HIV 1/2 3.0 and other CE marked, state-of-the-art rapid assay on 30 seroconversion panels were evaluated. When compared with EIA, the time delay of SD BIOLINE HIV 1/2 3.0
was 2 bleeds, 0.41 days and other CE marked rapid assay was 4 bleeds, 0.66 days.
*The classification of both seroconversion HIV samples and early HIV seroconversion samples is done according to the definition of each terms specified in the CTS 2009/886/EC for In Vitro Diagnostics Medical Device.
14 seroconversion HIV samples and 183 early HIV seroconversion samples classified among 30 seroconversion HIV panels were evaluated with SD BIOLINE HIV 1/2 and other CE marked, state-of-the-art rapid assay.
[Seroconversion HIV samples]
One step rapid test for the detection of antibodies to HIV-1/HIV-2 seroconversion HIV samples mean p24 antigen and/or HIV RNA positive, and recognized by all of the antibody screening tests, and positive or indeterminate confirmatory assays.
The term ‘rapid test’ is understood to mean those tests which can only be used singly or in a small series and which have been designed to give a rapid result for near patient testing. SD BIOLINE HIV 1/2 other CE marked HIV assay
seroconversion HIV samples(14) 14/14 (Recognized) 14/14 (Recognized)
Explanation of the test [Early seroconversion HIV samples]
[Introduction] HIV (Human Immunodeficiency Virus) is recognized as the etiologic agent of Aquired Immune Deficiency Syndrome (AIDS). The virus is transmitted by sexual contact, exposure to infected blood, certain body fluids Early seroconversion HIV samples mean p24 antigen and/or HIV RNA positive, and not recognized by all of the antibody screening tests, and indeterminate or negative confirmatory assays.
or tissues, and from mother to fetus or child during the perinatal period. HIV-1 has been isolated from patients with AIDS and AIDS related complex, and from healthy persons with high potential risk of developing AIDS. Patients SD BIOLINE HIV 1/2 other CE marked HIV assay
with HIV-2 are found primarily in parts of West Africa. Its course is marked by increasing levels of viral replication and the emergence of more virulent viral strains. This process causes the destruction of the immune system. HIV Early seroconversion HIV samples(183) 43/183 43/183
infection is staged by CD4 cell counts and clinical symptoms. Not all people progress through all "stages" and the time frames may also vary greatly from person to person.
HIV-1 and HIV-2 are similar in their morphology, cell tropism, host interaction and generic structure. Serological studies have determined that HIV-1 and HIV-2 have multiple common epitopes in core antigens but much less so 3. Diagnostic specificity
in the envelope antigens. This clinical diagnostic issues related to HIV are the detection of antibodies to HIV1/2 in human plasma or serum by immunoassay. Among the existing immunological method, the ELISA format and
Category No. of samples tested SD HIV 1/2 3.0
immunochromatographic format (rapid) to detect antibodies to HIV1/2 are available.
Synthetic peptides and recombinant antigen defining the antigenic determinants of HIV proteins find wide applications in ELISA procedure for the detection antibodies to HIV in serum. Recently, the use of recombinant antigen for Negative Positive
both detector and capture materials markedly increased both the sensitivity and the specificity compared to a commercial rapid kit based on synthetic peptides. Researchers of Standard Diagnostics, Inc. have constructed HIV1/2 Healthy donors 500(*3) 500 0
genes for the expression of recombinant antigens in bacterium systems such as E. coli and focused on HIV1/2 proteins, which are definitely immunogenic. The major immunoreactive antigens of these proteins have been reported 250(*4) 250 0
to have HIV-1 gp41, p24 and HIV-2 gp36 based on western blot analysis. It is known that 10 % of normal person has antibody against E.coli, Researchers of Standard Diagnostics, Inc. also tested cross reactivity with rabbit anti- E.coli. 250(*5) 250 0
in order to check the cross reactivity of human anti- E.coli. The test result was shown there is no cross reactivity of Rec. HIV1/2 Ag originated from E.coli and human anti-E.coli.
[Test principle] The SD BIOLINE HIV 1/2 3.0 kit is a rapid, qualitative test for the detection of antibodies to all isotypes (IgG, IgM, IgA) specific to HIV-1 including subtype-O and HIV-2 simultaneously in human serum, plasma Possible interferences (*2,6) 500 498 2
or whole blood. The SD BIOLINE HIV 1/2 3.0 test contains a membrane strip, which is precoated with recombinant HIV-1 capture antigen (gp41, p24) on test line 1 region and with recombinant HIV-2 capture antigen (gp36) on Total 1500 1498 (99.87%) 2
test line 2 region respectively. The recombinant HIV-1/2 antigen (gp41, p24 and gp36)-colloid gold conjugate and the sample move along the membrane chromatographically to the test region (T) and forms a visible line as the
antigen-antibody-antigen gold particle complex forms with high degree of sensitivity and specificity . The Test line and control line in the result window have been clearly label : “1” for test line 1 and “2” for test line 2 and “C” for 4. Possible interferences
Control line. Both test line and control lines in the result window are not visible before applying any sample. The Control line is used for procedural control and should always appear if the test procedure is performed correctly. The table below shows the results of SD BIOLINE HIV 1/2 tested on a variety of samples containing possibly interfering substances.
[Intended Use] The SD BIOLINE HIV 1/2 3.0 kit is a rapid, qualitative test for the detection of antibodies to all isotypes(IgG, IgM, IgA) specific to HIV-1 and HIV-2 simultaneously in human serum, plasma or whole blood. The SD
HIV type No. of samples tested SD HIV 1/2 3.0
BIOLINE HIV 1/2 3.0 kit is intended for professional use, only for an initial screening test and reactive samples should be confirmed by a supplemental assay such as ELISA or Western Blot test.
Negative Positive

Materials Provided / Active ingredients of main components Hospitalized patients (*2)

Pregnant women (*6)
200
200
198
200
2
0
1. The SD BIOLINE HIV 1/2 3.0 test kit contains the following items to perform the assay.
Potentially cross reacting samples*(*6) 100 100 0
• Test devices individually foil pouched with a desiccant
• Assay diluents * The results were negative for samples containing Anti-HBV(50), Anti-HCV(5), Anti-CMV antibody positive(20), Anti-EBV antibody positive(20), Rheumatoid factor positive(2), Anti-HTLV-I/HTLV-II positive(3)
• 20µl capillary pipettes(Option), Lancets (Option)
• Instructions for use 5. Mutant analysis(*2)
2. Active ingredients of main components The table below shows the results of SD BIOLINE HIV 1/2 and other CE marked HIV assay tested on 5 HIV-1 positive plasma samples (originating from patients infected with different HIV-1 env mutant viruses)
• 1 test strip included ; Gold conjugate (as main component) : Recombinant HIV-1 gp41, p24, HIV-2 gp36 antigen – gold colloid(1±0.2µg), Test line 1 (as main component) : Recombinant HIV -1 antigen (gp41, p24) Sample ID HIV-1 RNA copies/ml Mutated region analyzed SD HIV 1/2 3.0 other CE marked HIV assay
(0.625±0.125µg), Test line 2 (as main component) : Recombinant HIV -2 antigen (gp36) (0.5±0.1µg), Control line : Goat anti-HIV serum (0.75±0.15µg) (Nuclisens Easy Q)
• Assay diluents : 50mM Tris-HCl Buffer, Sodium azide(0.02w/v%) 3098/1988 160,000 env Positive Positive
3104/1986 6,700 env Positive Positive
Materials required but not provided 3107/1988 490,000 env Positive Positive
Micropipette 3108/1977 7,700 env Positive Positive
3109/1974 38,000 env Positive Positive
Precautions / Kit Storage and Stability
1. The test device should be stored at 1-30°C. Do not freeze the kit or components. 6. Equivalence studies(*2)
2. If assay diluents is kept at 1~30°C, it is stable until the expiration date after the first opening of the bottle. [Serum to Whole blood]
3. The test device is sensitive to humidity as well as to heat. Specimen type No. of SD BIOLINE neg./ No. of true neg. samples No. of SD BIOLINE pos./ No. of true pos. samples
4. Perform the test immediately after removing the test device from foil pouch. Serum 25/25 25/25
5. Do not use it beyond the expiration.
Venous blood 25/25 25/25
6. The shelf-life of the kits is as indicated on the outer package.
7. Do not use the test kit if the pouch is damaged or the seal is broken. Fingerstick blood 25/25 25/25
8. Check the desiccant for color change of the humidity indicator and to throw away the pouch if the color indicates saturation. [Serum to Plasma(each anticoagulant)]
9. Do not re-use the test device. Specimen type No. of SD BIOLINE neg./ No. of true neg. samples No. of SD BIOLINE pos./ No. of true pos. samples
Serum 25/25 25/25
Warnings & Precautions Plasma(heparin) 25/25 25/25
[Precautions under the HIV regulations] Plasma(EDTA) 25/25 25/25
1. Negative result may not have detected recently acquired HIV infection. Plasma(citrate) 25/25 25/25
[Safety precautions] * All samples are “same day” fresh samples (≤1day after sampling).
1. Wear protective gloves while handling specimens. Wash hand thoroughly afterward.
2. Do not pipette by mouth. 7. Precision(*7)
3. Do not eat or smoke while handling specimens. • Within-run: the reproducibility was determined by SD’s laboratory personnel testing 5 different replicates of 9 different specimens containing different concentration of antibody. All qualitative results obtained were
4. Clean up spills thoroughly using an appropriate disinfectant. 100% in agreement with the expected results.
5. Decontaminate and dispose of all specimens, reaction kits and potentially contaminated materials, as if they were infectious waste, in a biohazard container. • Between-run: the reproducibility was determined by three SD’s laboratory people testing in replicates of 9 different specimens containing different concentration of antibody. All qualitative results obtained were 100%
6. SD BIOLINE HIV 1/2 3.0 assay diluents contain a proprietary anti-microbial agent which presents no harzard to the user if normal laboratory safety precautions are followed. in agreement with the expected results.
[Handling precautions] • Batch-to-batch performance: the reproducibility was determined by testing in replicates of 9 different specimens containing different concentration of antibody with three different lots. All qualitative results
1. Avoid splashing or aerosol formation. obtained were 100% in agreement with the expected results.
2. Do not mix and interchange different specimen.
*Testing sites 1 : Testing Laboratory for In Vitro Diagnostic Devices at the Paul Ehrlich Institute
3. Serum or plasma specimens may be stored for 2 weeks at 2-8°C before testing.
4. Anticoagulants such as heparin, EDTA and sodium citrate do not affect the test result. 2. : National Center for Epidemiology Microbiological Research Group
5. Use of hemolytic samples, rheumatoid factors-contained samples and lipidemic, icteric samples can lead to impair the test result. 3. : German Red Cross, Hessen
[Warnings] 4. : National Blood Bank, Regional Center, Buda, Budapest
1. For in vitro diagnostic use only. 5. : National Health Center, Department of Transfusion and Diagnostics, Budapest
2. The instruction must be followed exactly to get accurate results. Anyone performing an assay with this product must be trained in its use and must be experienced in laboratory procedures. 6. : National Center for Epidemiology, division of Virology, Budapest
3. Care should be taken to avoid contamination of the end of bottle when dropping of assay diluents into sample well. 7. : SD Biotech Center Laboratory, Republic of Korea.

Specimen collection, storage and precaution Bibliography of suggested reading

Whole blood
1. McCutchan, F.E. (2006). Global epidemiology of HIV. J. Med. Virol. 78, Suppl. 1, S7-S12.
[Collection by venipuncture]
2. Branson, B.M. (2003). Point-of Care Rapid Tests for HIV Antibodies. J. Lab. Med. 27, 288-295.
• Collect the whole blood into the collection tube (containing anticoagulants such as heparin, EDTA and sodium citrate) by venipuncture.
3. Ferreira Junior, O.C., Ferreira, C., Riedel, M., Widolin, M.G.V., and Barbosa-Junior, A. (2005). Evaluation of Rapid tests for anti-HIV detection in Brazil. AIDS 19 (suppl 4), S70-S75.
• If blood specimens are not immediately tested, they should be refrigerated at 2~8°C.
4. Constantine, N. (2006). HIV Antibody Assays. HIV InSite Knowledge Base Chapter.
• When stored at 2~8°C, the blood specimens should be used within 3 days.
5. Castilla, J., Pachon, I., Gonzalez, M.P., Amela, C., Munoz, L., Tello, O., Noguer, I., de Ory, F., Leon, P., Alonso, M., Gil, E., and Garcia- Saiz, A. (2000). Seroprevalence of HIV and HTLV in a representative sample of the Spanish population. Epidemiol.
• Using the blood specimens in the long-term keeping more than 3 days can cause nonspecific reaction.
Infect. 125, 159-162.
[Collection using a lancet]
6. Manfredi, R., Calza, L., and Chiodo, F. (2001). HIV disease among immingrants coming to Italy from outside the European Union: a case-control study of epidemiological and clinical features. Epidemiol. Infect. 127, 527-533.
• Clean the area to be lanced with an alcohol swab.
7. Lot, F., Semaille, C., Cazein, F., Barin, F., Pinget, R., Pillonel, J., and Desenclos, J.C. (2004). Preliminary results from the new HIV surveillance system in France. Eurosurveillance 9, 10-12.
• Squeeze the end of the fingertip and pierce with a sterile lancet provided.
8. Soriano, V., Gomes, P., Heneine, W., Holguin, A., Doruana, M., Antunes, R., Mansinho, K., Switzer, W.M., Araujo, C., Shanmugam, V., Lourenco, H., Bonzalez-Lahoz, J., Antunes, F. (2000). Human immunodeficiency virus type 2 (HIV-2) in Portugal:
• Take a 20μl capillary pipette provided, immerse the open end in the blood drop and then release the pressure to draw blood into the capillary pipette to black line.
clinical spectrum, circulating subtypes, virus isolation, and plasma viral load. J. Med. Virol. 61, 111-116.
Plasma or Serum
• [Plasma] Collect the whole blood into the collection tube (containing anticoagulants such as heparin, EDTA and sodium citrate) by venipuncture and then centrifuge blood to get plasma specimen.
In vitro diagnostic medical device Use by : Exp Batch code
• [Serum] Collect the whole blood into the collection tube (NOT containing anticoagulants such as heparin, EDTA and sodium citrate) by venipuncture, leave to settle for 30 minutes for blood coagulation and then
centrifuge blood to get serum specimen of supernatant.
• If plasma or serum specimens are not tested immediately, they should be refrigerated at 2~8°C. For storage period longer than 2 weeks, freezing is recommended. They should be brought to room temperature prior to
Do not re-use Authorized representative in the european Catalogue number
use.
community
• Plasma or serum specimens containing a precipitate may yield inconsistent test results. Such specimens must be clarified prior to assaying.
Consult instructions for use Manufacturer Contains sufficient for <n> tests
Procedure of the test (Refer to figure) n
1. Remove the test device from foil pouch, place it on a flat, dry surface. Temperature limitation
2. [Using a capillary pipette]
Add 20µl of drawn blood specimen with a 20µl capillary pipette into the sample well(s).
OR,
[Using a micropipette]

3.
Add 10µl of plasma or serum specimen (20µl of blood specimen) into the sample well(s).
Add 4 drops of assay diluents vertically into sample well(s).
Information about usage of the lancets provided
Caution : If you do not hold the bottle vertically, it can lead to inaccurate results. Exactly, 4 drops should be added. Adding 5 or 6 drops (i.e. more than 4 drops) may cause reverse migration phenomenon and/or overall a 1. Intended Use : 3. Warning / Precautions :
little reddish unclear background due to limitation of absorbent pad capability. Lancet is intended for skin puncture to get a drop of blood from the fingertip. For single use only. It is • DO NOT share used lancets with another person. To prevent possible infection, a used lancet
4. As the test begins to work, you will see purple color move across the result window in the center of the test device. sterilized by EO gas sterilization. For professional use only. should not be touched by another person.
5. Time to result is 10 to 20 minutes. After adding the diluent, read the result after 10 minutes but not more than 20 minutes. • DO NOT use expired lancet. The use of an expired lancet may cause any infection at the punctured
Caution : If the test result is not legible after 10 minutes due to high background color, read again later but within 20 minutes of adding the diluent. Do not read after 20 minutes. 2. Test Procedure : skin due to cease to exist its sterility
• Massaging the fingertip gently will help you obtain a • Store at room temperature (0 ~ 40 °C)
Interpretation of the test (Refer to figure) round drop of blood.
1. A color band will appear in the left section of the result window to show that the test is working properly. This band is control line (C). • Twist off the protective cap of lancet.
2. Color bands will appear in the middle and right section of the result window. These bands are test line 2 and test line 1 (2, 1). • Squeeze the end of the fingertip and pierce with a
Negative Result lancet provided.
• The presence of only control line (C) within the result window indicates a negative result. • Gently squeeze the area until you get a small sample of
Positive Result blood. And take a 20 µl capillary pipette provided.
• The presence of two lines as control line (C) and test line 1 (1) within the result window indicates a positive result for HIV-1. Note: A lancet should only be used once. Dispose of
• The presence of two lines as control line (C) and test line 2 (2) within the result window indicates a positive result for HIV-2. used lancets in accordance with local regulatory
• The presence of three lines as control line (C), test line 1 (1) and test line 2 (2) within the result window indicates a positive result for HIV-1 and/or HIV-2. guidelines, and in a safe manner so as not to cause accidental injury.
- If the color intensity of the test line 1 is darker than one of test line 2 in the result window, you can interpret the result as HIV-1 positive.
- If the color intensity of the test line 2 is darker than one of test line 1 in the result window, you can interpret the result as HIV-2 positive.
Caution : Although a positive result for HIV-1 and HIV-2 in one patient is a rare case, it's possible as there is an homology in the amino acid sequence between HIV-1 and HIV-2. To determine the virus type or diagnose a co-
infection accurately, you must perform a confirmatory test as Western Blot etc. Date issued : 2013. 09
Invalid Result 03FK10CE-En-3 / 03FK16CE-En-2 / 03FK18CE-En-2 / 03FK10LCE-En-2
• No presence of control line (C) within the result window indicates an invalid result. The directions may not have been followed correctly or the test may have deteriorated. It is recommended that the specimen be re-
tested.

Limitations of the test Manufactured by

1. Although a positive result may indicate infection with HIV-1 or HIV-2 virus, a diagnosis of AIDS can only be made on clinical grounds, if an individual meets the case definition for AIDS established by the Centers fir
Disease Control. For samples repeatedly tested as positive, more specific supplemental tests must be performed.
2. A negative results does not eliminate the possibility of HIV-1 / HIV-2 infection. The specimen may contain low levels of antibodies to HIV-1 / HIV-2. 65, Borahagal-ro, Giheung-gu, Yongin-si, Gyeonggi-do, Republic of Korea
3. Specimens repeatedly tested positive should be retested using another method. Tel : 82-31-899-2800 Fax : 82-31-899-2840
4. Immunochomatographic testing alone cannot be used to diagnose AIDS even if the antibodies against HIV-1 and/or HIV-2 are present in a patient specimen. http://www.standardia.com sales@standardia.com
5. As with all diagnostic tests, a definitive clinical diagnosis should not be based on the results of a single test, but should only be made by the physician after all clinical and laboratory findings have been evaluated. Shandong
6. The whole blood application on seroconversion panels could not be evaluated due to unavailability of such panels made up of whole blood specimens. No.1 Shuangshan Sanjian Road 250200 Zhangqiu City, Jinan,
Authorized Representative
Shandong PEOPLE’S REPUBLIC OF CHINA
Internal Quality Control Shanghai
Altenhofstrasse 80 D-66386 St. Ingbert Germany
The SD BIOLINE HIV-1/2 3.0 test device incorporates the characters “1”, “2” and “C” on the surface of the device to identify the adjacent location of the Test Line (HIV-1), Test Line (HIV-2) and Control Line in the result window.
These lines are not visible before applying the sample. The Control Line is used for procedural control. A visible Control Line confirms that the diluents has been applied successfully and that the active ingredients on the strip are
Phone : +49 6894 581020, Fax : +49 6894 581021
functional, but is not confirmation that the sample has been properly applied; it is not a positive sample control.
 The 3rd Generation of ONE STEP antibodies to HIV-1/HIV-2 Test I. Blood specimen (with a lancet) Interpretation
Time to result is 10 to 20 minutes. After adding the diluent, read the result after 10 minutes but not more than

4 5 6
20 minutes. If the test result is not legible after 10 minutes due to high background color, read again later but
within 20 minutes of adding the diluent. Do not read after 20 minutes.

Test Procedure How to interpret test results

Bla
Blo
od
ck L
ine Negative

1
Now, open the package and look for the following :
1. Test devices individually foil pouched with a desiccant
Clean the area to be lanced with an
Positive
2. Assay diluents Pierce with a sterile lancet provided. Take a 20µl capillary pipette provided, immerse the open end
3. Instructions for use alcohol swab. in the blood drop and then release the pressure to draw blood
into the capillary pipette to black line.
HIV-1 Positive
2 lines 3 lines
1 2 3 When 2 lines appear When 3 lines appear
ONE STEP Anti-HIV 1/2 TEST 1 Test "C", "1" Line "C","1" and faint "2" Line

7
HIV-1/2 3.0

8
03FK10CE-P-1

30°C
1°C
For in vitro diagnostic use only.
Store at 1~30ºC sealed.

Strong

Option
1. 20μl capillary Pipettes Medium
2. Lancets
1 2 20μl 4 drops If line "1" is darker than line "2", it should be interpreted as HIV-1 positive only.
( Not HIV- 2 positive ).
Faint line : By homology in the amino acid sequence of HIV Type-1and HIV Type-2.
Dispense Weak

Open the foil pouch and look for the following.

HIV-2 Positive
Add 20µl of drawn blood into the sample well (s). Add 4 drops of assay diluents into the sample well (s).
Result window Exactly, 4 drops should be added. 2 lines 3 lines
When 2 lines appear When 3 lines appear
ONE STEP Anti-HIV 1/2 TEST 1 Test
"C", "2" Line "C", "2" and faint "1" Line
HIV-1/2 3.0
먹지 마세요

II. Blood (by venipuncture), Plasma or Serum specimen

03FK10CE-P-1

Valid Invalid
30°C
1°C
For in vitro diagnostic use only.
Store at 1~30ºC sealed.

Strong
Sample well & Assay diluents well
4 5
2
F IRST, read carefully the instruction on how to use the SD BIOLINE HIV-1/2 3.0 test
Medium

kit. If line "2" is darker than line "1", it should be interpreted as HIV-2 positive only.
( Not HIV- 1 positive ).
Weak Faint line : By homology in the amino acid sequence of HIV Type-1and HIV Type-2.

4 drops
Remark
3
Next, look at the expiry date at the back of the foil pouch.
10μl (Plasma or Serum)
or 20μl (Blood)
[Both HIV 1 and HIV 2 Positive]
If the band thickness between line 1 and line 2 is so similar to each other , even very rare , it can be both positive for HIV-1 and HIV- 2. In this case, confirm test using Western Blot
is recommended to know exact virus type.
Use another kit, if expiry date has passed.
Invalid
[ For example ]
Add 10µl of plasma or serum (20µl of blood) into the sample well (s) using a Add 4 drops of assay diluents into the sample well (s). No"C" line
××××××
micropipette. Exactly, 4 drops should be added.
××××××
××××. ××. ××
××××. ××. ××
××××. ××. ××