New Product Launch - bioelisa HIV 1+2 Ag/Ab

NEWS BK NEWS# 359 / BE/ MKT DATE : December 19, 2014 TITLE : New Product Launch - bioelisa HIV 1+2 Ag/Ab Dear customer, With the aim of continuous development of its products, Biokit now presents a new member of the retrovirus family: The bioelisa HIV 1+2 Ag/Ab is a fourth generation Elisa test for the detection of antibodies to HIV 1+2 and the HIV-1 p24 Antigen. The available presentations are the following: bioelisa HIV 1+2 Ag/Ab 96 Tests, Cod. 3000-1172 480 Tests, Cod. 3000-1173 The bioelisa HIV 1+2 Ag/Ab includes new features that improve the performance of the existing bioelisa HIV 1+2 4.0 assay. - New HIV-1 gp41 recombinant antigen for better specificity - Two monoclonal antibodies for better sensitivity to HIV-1 p4ag - Two kit presentations for better adjustment to customer needs According to the internal and external evaluations, the new assay overcomes the performance of the today s state-of-the art HIV assays: Biorad Genscreen HIV1+2 Ultra and Abbott Architect HIV1+2 Combo The bioelisa HIV 1+2 Ag/Ab will substitute the bioelisa HIV 1+2 4.0. It is important to point out that the bioelisa HIV 1+2 4.0 will be available in Biokit during the time needed for registering the bioelisa HIV 1+2 Ag/Ab in the non-european Union countries. Public tenders already awarded with the bioelisa HIV 1+2 4.0 will be supplied without restrictions. Please find enclosed a complete product information that we hope will help you in the promotion and introduction of bioelisa HIV 1+2 Ag/Ab.

bioelisa HIV 1+2 Ag/Ab Product features: Recombinant antigens and capture antibody bioelisa HIV 1+2 Ag/Ab uses a combination of two recombinant antigens: gp41 from HIV-1 and gp36 from HIV-2, in addition to a synthetic peptide for HIV-1 subtype O, together with a two monoclonal antibodies against HIV-1 p24 Ag. The new HIV antigens are able to detect HIV type 1 group M-O and HIV type 2 in human serum or plasma. Performance of bioelisa HIV 1+2 Ag/Ab in terms of sensitivity and specificity allowed biokit to achieve the Common Technical Specifications (CTS) requested for CE mark class IIA products. Fourth generation assay The bioelisa HIV 1+2 Ag/Ab is a fourth generation assay for total antibody detection and HIV-1 p24 antigen detection. HIV recombinant antigens and synthetic peptides are present in both solid phase and conjugate. If HIV antibodies (IgG, IgM or IgA) are present in the patient sample, they will bind to the HIV antigens on the plate coating and HIV antigens present in the conjugate. If p24 Antigen is present in the patient sample, it will bind to the capture monoclonal antibody present in the plate coating and capture antibody present in the sample diluent. Components MICROPLATE: 12 strips of 8 breakable wells coated with HIV specific gp36 and gp41 peptides and gp41 protein with two monoclonal antibodies specific to the HIV-1 p24 Ag. NEGATIVE CONTROL Ready-to-use control. It contains human serum negative for HIV antibodies and p24 antigen. POSITIVE HIV-1 CONTROL Ready-to-use control. It contains human serum positive to Anti- HIV 1. POSITIVE HIV-2 CONTROL Ready-to-use control. It contains human serum positive to Anti- HIV 2. POSITIVE HIV p24 CONTROL Ready-to-use control. It contains recombinant HIV-1 p24. CONJUGATE #1: Ready-to-use, blue colour-coded reagent. It contains a mixture of biotinylated antigens in phosphate buffer. CONJUGATE #2 CONC. 100x: 100x solution concentrate. It contains Horseradish Peroxidase conjugated with streptavidin in specific stabilizer reagent to be diluted with Conjugate # 2 Diluent. Page 2 of 15

CONJUGATE #2 DILUENT: Ready-to-use orange colour-coded reagent used for the dilution of Conjugate # 2 Concentrate 100x. SAMPLE DILUENT: Ready-to-use red-colour coded reagent. It contains active components and monoclonal p24 antigen antibodies. Used to dilute the sample. SUBSTRATE TMB: Ready-to-use component. It contains Tetra methyl benzidine and Hydrogen peroxide (H2O2) in a citrate buffer. STOP SOLUTION ready-to-use component. It contains 0,3 M H2SO4 solution. 25x concentrated solution. WASH BUFFER CONC. 25x: 25x concentrated solution. It contains saline phosphate buffer. ADHESIVE SEALS: To cover the microplate during incubation. Protocol: The bioelisa HIV 1+2 Ag/Ab uses in plate sample dilution: 100 µl of sample diluent and 100 µl of sample or control. The 1:2 sample dilution allows spectrophotometric monitoring of sample dispensing at 620 nm. The red colour of the sample diluent changes to a clear red dark colour. Definition of the OD limit must be validated customer by customer. Number of replicates controls have been reduced to allow the maximum well utilization: Negative control by triplicate and rest of Positive control for HIV-1, HIV-2 and p24 Ag, only by singlicate. The sample incubation time is to 60 minutes to reach the required sensitivity of fourth generation HIV assays. Conjugate incubation is split into two separate incubations of 30 minutes each because of the biotin streptavidin system. The substrate incubation is 30 minutes. The total incubation time is 2 hours and 30 minutes. Substrate is ready for use. STEP PROCEDURE Add 100 µl of Sample Diluent. Add 100 µl of sample/control to the diluent and pipette up & down for Sample step mixing. Test Negative control x3 Positive Controls x1. Incubate 60 minutes at 37 C. Wash step Perform wash step 5x. Conjugate #1 Add 200 µl of ready-to-use Conjugate #1 to wells of the microtiter plate. Incubate 30 minutes at 37 C. Wash step Perform wash step 3x. Conjugate #2 Add 200 µl of diluted Conjugate #2 to wells of the microtiter plate. Incubate 30 minutes at 37 C Wash step Perform wash step 5x. TMB Substrate Add 150 µl of TMB Substrate to the wells of the microtiter plate. Colour Development Incubate 30 minutes at 18-25 C. Stopping Add 100 µl of Stop Solution and read OD at 450nm with reference wavelength at 600-650nm in an ELISA microplate reader. Page 3 of 15

Comparison with bioelisa HIV 1+2 Ag/Ab: Test name bioelisa HIV 1+2 4.0 bioelisa HIV 1+2 Ag/Ab Code number 3000-1175 192 Tests 3000-1172 96 Tests 3000-1173 480 Tests Method EIA/HRP/TMB EIA/HRP/TMB Format Solid phase 3 - step direct sandwich 12x8 Microtiter wells 3 - step direct sandwich 12x8 Microtiter wells Coating Recombinant antigens and synthetic peptides Recombinant antigens and synthetic peptides HIV-1 gp41 gp41 HIV-2 gp36 gp36 Anti p24 Ag One monoclonal antibody Two monoclonal antibodies Conjugate Substrate Specimen dilution Number of controls Performance: Conjugate #1: Biotinylated HIV Ag Ready-to-use Conjugate #2: Streptavidin HRP Concentrate Ready-to-use 150 µl 100 µl Sample diluent 100 µl Sample Blk, NC (x2), HIV1(x2), HIV2(x2), p24(x2): 9 wells in total Conjugate #1: Biotinylated HIV Ag Readyto-use Conjugate #2: Streptavidin HRP Concentrate Ready-to-use 150 µl 100 µl Sample diluent 100 µl Sample Blk, NC(x3), HIV1(x1), HIV2(x1), p24(x1) 7 wells in total Sample incubation 60 min / 37 C 60 min / 37 C Conjugate incubation Conjugate #1: 30 min / 37 C Conjugate #2: 30 min / 37 C Conjugate #1: 30 min / 37 C Conjugate #2: 30 min / 37 C Substrate incubation 30 min / RT 30 min / RT Cut-off Mean Neg. + 0.170 Mean Neg. + 0.170 Diagnostic Sensitivity Analytical Sensitivity 100% 1 IU/ml 100% 0.5 IU/ml Specificity 99.7 99.87 Comments Fourth generation Nine wells per run One Kit presentation: 192 Tests Fourth generation Improved specificity and analytical sensitivity Only seven wells per run Two Kits presentation: 96/480Tests Sensitivity: Analytical Sensitivity (Limit of Detection HIV-1 p24 Ag) The analytical sensitivity of the test was determined using WHO International Standard HIV-1 p24 Antigen and the BIORAD HIV-1 Ag Standard. The WHO International Standard HIV-1 p24 Antigen and the BIORAD HIV-1 Ag Standard were serially diluted in negative human serum and analysed in duplicate with the test assay. Tables are showing the different dilutions of the HIV-1 Antigen standards, the absorbance results, and ABS/CO values have been prepared, from two different lots. The analytical sensitivity was determined as the concentration of the analyte in the highest dilution of the standards yielding an ABS/CO > 1.0. Lot 1 Lot 2 WHO Int std HIV-1 p24 OD/COV 2 IU/ml 5.02 5.61 1 IU/ml 2.64 2.83 0.5 IU/ml 1.44 1.49 End point 0.32 IU/ml 0.32 IU/ml The sensitivity limit has been estimated at < 0.5 IU/mL Page 4 of 15

Analytical sensitivity AFSSAPS/Biorad standard Lot 1 Lot 2 Biorad HIV-1 Ag std OD/COV 100 pg/ml 8.89 9.17 50 pg/ml 4.68 4.85 25 pg/ml 2.62 2.72 12.5 pg/ml 1.40 1.53 8.3 pg/ml 1.00 1.04 End point 8.3 pg/ml 8.3 pg/ml The sensitivity limit has been estimated at < 12.5pg/ml The analytical sensitivity for antibody detection was determined using the NIBSC International Standards: British Working Standard HIV-1, British Working Standard HIV-1 1 in 5 dilution and Monitor Sample for HIV-2. Lot 1 Lot 2 OD/COV British Working Std HIV-1 7.54 6.38 British Working Std HIV-1 diluted 1:5 1.93 1.60 Monitor Sample HIV-2 8.71 9.62 The HIV-1 p24 Ag diagnostic sensitivity was assessed by using the Seracare HIV p24 Antigen Mixed Titer Performance panel PRA204 All samples in this panel are positive for the presence of HIV-1 p24 Ag. In the next graphic it is showed the reactivity in terms of signal Page 5 of 15

OD/COV for the most difficult samples in comparison with other commercial assays. The bioelisa HIV 1+2 Ag/Ab presented the best score in all of them. Diagnostic Sensitivity The diagnostic sensitivity of the bioelisa HIV 1+2 Ag/Ab Kit was based on testing characterized HIV-1, HIV-2 and HIV-1 p24 Ag positive samples: - A total of 537 HIV-1 positive samples were evaluated with bioelisa HIV 1+2 Ag/Ab, among them: 28 HIV-1 fresh-same-day collected samples, 85 HIV-1 and 4 HIV-1 Subtype O samples in an European reference lab; 380 HIV -1 samples were evaluated at Biokit and 40 HIV-1 non B subtype samples (and at least 3 samples of each HIV-1 subtype) were evaluated in another European reference lab. bioelisa HIV 1+2 Ag/Ab detected all 537 samples. The sensitivity found was100% (537/537). - A total of 115 HIV-2 positive samples were evaluated with bioelisa HIV 1+2 Ag/Ab, among them: 27 HIV-2 samples in an European reference lab and 88 HIV-2 samples were evaluated at Biokit. bioelisa HIV 1+2 Ag/Ab detected all 115 samples. The sensitivity found was 100% (115/115). - A total of 61 HIV-1 p24 Ag positive samples were evaluated also with bioelisa HIV 1+2 Ag/Ab, among them: 15 samples in an European reference lab, 16 samples were evaluated in another European reference lab and 30 samples at Biokit. bioelisa HIV 1+2 Ag/Ab detected all 61 samples. The sensitivity found was 100% (61/61). - Additional sensitivity studies were carried out by testing a total of 31 seroconversion panels at Biokit and a European reference lab. Results obtained were comparable to the most sensitive commercial method for HIV 1+2 antigen and antibody detection. Page 6 of 15

- 16 HIV Seroconversion Panels from BBI/Seracare and Zeptometrix were evaluated at Biokit. bioelisa HIV-1+2 Ag/Ab identified correctly all the positive specimens. In the following table it can be observed the first sample detected as positive in each panel evaluated with bioelisa HIV-1+2 Ag/Ab in comparison with 4 th Generation assays (Ag/Ab Assays), 3 rd Generation assays (only Ab Assays), assays for HIV p24 antigen and NAT HIV RNA assays. bioelisa HIV-1+2 Ag/Ab showed excellent sensitivity to detect early antigen or antibodies in 16 HIV Seroconversion Panels evaluated. bioelisa HIV Ag/Ab HIV p24 Seroconversion HIV Ab Assay NAT HIV Ag/Ab Assay antigen Panel First sample detected positive in the panel BB1-PRB965-n=6 2 2 4 2 1 ZEPT-12007-n=9 4 4 5 ND 4 BBI-PRB954-n=7 6 6 7 6 4 BBI-PRB932-n=9 4 ND 4 4 4 ZEPT-9017-n=11 1 4 3 1 1 ZEPT-9014-n=7 1 1 3 1 1 ZEPT-9012-n=8 5 6 7 5 1 ZEPT-9032-n=14 8 8 7 8 1 ZEPT-6243-n=10 7 7 8 7 1 ZEPT-9022-n=9 7 8 9 7 1 ZEPT-9018-n=11 9 9 10 9 1 BBI-PRB939-n=9 6 ND 8 6 5 BBI-PRB956-n=5 4 4 5 4 2 BBI-PRB945-n=6 3 4 4 3 1 BBI-PRB955-n=4 2 2 4 2 1 BBI-PRB958-n=6 3 3 5 3 1 Bioelisa HIV-1+2 Ag/Ab detected the first positive sample at the same level as an Elisa for only HIV p24 detection and before than a commercial EIA assay for HIV1+2 Ag/Ab detection In the next graphic it is shown the aggregate score in seven seroconversion panels, in comparison with other commercial assays. The aggregate score means the addition of the first detection day in all panels. A lower aggregate value means earlier detection and higher sensitivity. Page 7 of 15

Maximum number detection days by less sensitive HIV CE mark 3rd generation assay: #238. bioelisa HIV 1+2 Ag/Ab showed the best performance. - Another study was conducted to assess the sensitivity in 32 early seroconversion samples. These type of samples are characterized to be positive to the presence of HIVp24 Ag and/or positive by NAT (Nucleic Acid Testing), not detected by all HIV 3rd generation methods and to be negative or Indeterminate by Western-Blot methods. bioelisa HIV 1+2 Ag/Ab detected all 32 samples. The sensitivity found was 100% (32/32). - 50 cell culture supernatans from HIV cell cultures, including different HIV-1 subtypes and HIV-2 were evaluated in a European Reference Lab. bioelisa HIV 1+2 Ag/Ab detected all 50 samples. The sensitivity found was100% (50/50). - Finally a study was conducted in the Paul Ehrilch Institute in Germany, to assess the sensitivity of bioelisa HIV 1+2 Ag/Ab by comparing reactivity with other commercial assays in 10 seroconversion panels. bioelisa HIV 1+2 Ag/Ab showed the best performance. Maximum number detection days by less sensitive HIV CE mark 3rd generation assay: #471 Specificity The specificity was evaluated by testing a total of 6,465 unselected blood donors' samples at three different sites. - In the first site 3,045 unselected samples including 684 first donors were tested. From this total, 4 samples were reactive. The specificity obtained in this study was 99.87% (3,041/3,045). - In the second site 500 unselected samples from Blood Bank were tested. All 500 samples were negative. The specificity was 100% (500/500) Page 8 of 15

- An internal evaluation of 2,920 unselected samples from Blood Bank was performed. From this evaluation, 6 samples were reactive. The specificity obtained in this study was 99.79%. (2,914/2,920) The final specificity was found to be 6,455/6,465: 99.85% Evaluation 1 (3,045 sera) Initial Reactive Repeated Reactive False Positive Specificity % Serum 6 4 4 99.87 Evaluation 2 (500 sera) Initial Reactive Repeated Reactive False Positive Specificity % Serum 0 0 0 100 Internal Evaluation (1,000 sera + 1,920 plasmas) Initial Reactive Repeated Reactive False Positive Specificity % Plasma 8 6 6 99.69 Serum 1 0 0 100 Total 9 6 6 99.79 - In addition 200 samples from hospitalized patients have been tested and compared to a reference test. 194 samples were found negative by both tests and six samples were reactive. The six samples were also reactive with other ELISA tests. A specificity of 100% was obtained in this study (194/194). Precision Intra-assay reproducibility: The coefficient of variation obtained for the absorbance values of a HIV-1 positive sample assayed in a minimum of 40 replicates was 3.42%, 5.79% and 4.71% in three lots studied. The coefficient of variation obtained for the absorbance values of a HIV-2 positive sample assayed in a minimum of 40 replicates was 6.39%, 5.91% and 8.38% in three lots studied. The coefficient of variation obtained for the absorbance values of a HIV-1 p24 positive sample assayed in a minimum of 40 replicates was 4.08%, 3.44% and 3.71% in three lots studied. Inter-assay reproducibility: Four positive samples of different levels were tested in 15 different assays. The coefficients of variation obtained for the ratios absorbance/cut-off of the four samples were 4.87%, 5.37%, 7.49% and 7.54%. Interferences Interference by addition No interference has been found for haemoglobin (500 mg/dl), bilirubin (20 mg/dl), human Albumin (5 g/dl) and triglycerides (500 mg/dl). Page 9 of 15

Cross-reactivity To evaluate possible interferences, 125 potential cross-reacting specimens were analyzed. Among those samples, 4 samples (RF+), 4 samples positive for anti-nuclear antibodies (ANA), 5 samples Mononucleosis, 89 samples from other related infectious diseases and 23 samples from pregnant women. No evidence of cross reactivity was found in the samples evaluated. The 2 anti-toxoplasma IgM samples that tested positive were also positive by other HIV assays. Potential interfering samples = 125 RF (Rheumatoid factor) - 4 Elevated IgG - 4 ANA (anti-nuclear Antibodies) - 4 Elevated IgM - 5 Mononucleosis - 5 HSV 1 IgG (Herpes Simplex Virus) - 5 Pregnant women- 23 (including 7 multiparous) HSV 2 IgG (Herpes Simplex Virus) - 4 Syphilis Ab - 6 anti-cmv IgG (Cytomegalovirus) - 5 anti-ebv (Epstein-Barr Virus) - 3 anti-cmv IgM (Cytomegalovirus) - 7 anti-rubella (Rubella Virus) - 4 anti-htlv (Human T-lymphotropic Virus) - 4 anti-vzv IgG (Varicela Zoster Virus) - 2 anti-hcv (Hepatitis C Virus) - 8 anti-vzv IgM (Varicela Zoster Virus) - 2 anti-e.coli (Escherichia coli) - 5 anti-toxoplasma IgG (T. gondii) - 10 HBsAg (Hepatitis B Antigen) - 8 anti-toxoplasma IgM (T. gondii) - 7 Comparison vs. Competitors bioelisa HIV 1+2 Ag/Ab can be compared with the two Premium products on the market: The Biorad Genscreen ULTRA HIV and the Murex HIV Ag/Ab combination. The sensitivity and specificity of bioelisa HIV 1+2 Ag/Ab matches what until now was considered the reference assay in Elisa, the Biorad Genscreen assay. The biokit assay is even better in terms of analytical sensitivity or limit of detection of HIV-1 p24 Antigen. Specificity is also better because of the utilization of an improved gp41 protein. In terms of ease-of-use, the bioelisa HIV 1+2 Ag/Ab incorporates all liquid reagents, in comparison with the Biorad and Murex assays that use lyophilized conjugates. There is a unique disadvantage compared to the Biorad and Murex assay. The bioelisa HIV 1+2 Ag/Ab uses two independent incubations for the conjugate incubation step. This means the total incubation time increases by 30 minutes, but the specificity and background of the assay improves because of this extra wash cycle. The internal and external evaluations performed on the bioelisa HIV 1+2 Ag/Ab kit demonstrate better performance with seroconversion panels and the detection limit for p24 Ag. Page 10 of 15

Competitor s table Test name bioelisa HIV 1+2 Ag/Ab Genscreen ULTRA HIV Murex HIV Ag/Ab Ag/Ab Combination Code number 3000-1172 72386 7G79-01 Method EIA/HRP/TMB EIA/HRP/TMB EIA/HRP/TMB Format 3 - step direct sandwich 2 - step direct sandwich 2 - step direct sandwich Solid phase 12x8 Microtiter wells 12x8 Microtiter wells 12x8 Microtiter wells Tests per kit 192 96/480 96/480 Coating Recombinant antigens Synthetic peptide Recombinant antigens Synthetic peptide Recombinant antigens Synthetic peptide HIV-1 gp41 Rec gp160 and Synthetic peptide HIV Group O ENV rec Ag and Synthetic peptide HIV Group O HIV-2 gp36 Synthetic peptide HIV2 ENV rec Ag Anti p24 Ag Two Monoclonal antibody One Monoclonal antibody One Monoclonal antibody Conjugate Conjugate #1: Biotinylated Conjugate #1: Biotinylated HIV Ag Ready to-use polyclonal Ab to p24 Ready to-use Conjugate #2: Streptavidin HRP Concentrate Conclusion Conjugate #2: Antigens labelled with lyophilized Streptavidin HRP A+B 80 µl 25 µl Conjugate #1 75 µl Sample Conjugate: Antigens and monoclonal antibodies conjugated with lyophilized HRP. Substrate Ready to-use 150 µl Ready to-use Specimen dilution 100 µl Sample diluent 25 µl Sample diluent 100 µl Sample 100 µl Sample Sample incubation 60 min / 37 C 60 min / 37 C 60 min / 37 C Conjugate incubation Conjugate #1: 30 min / 37 C Conjugate #2: 30 min / 37 C Conjugate: 30 min / 37 C Conjugate #2: 30 min / 37 C Substrate incubation 30 min / RT 30 min / RT 30 min / 37 C Cut-off Mean Neg. + 0.170 Mean Neg. + 0.200 Mean Neg. + 0.150 Analytical sensitivity 0.5 IU/ml p24 Ag 0.85 IU/ml p24 Ag 28 pg/ml Biorad std. 12.5 pg/ml Biorad std. 13.6 pg/ml Biorad std Diagnostic Sensitivity 100% 100% 100% Specificity 99.87% 99.72% 99.78% Comments + Best analytical sensitivity + 2h incubation time +Reference assay in the + Best sensitivity - Lyophilized conjugate market for diagnostic + Very low background sensitivity - 2h 30 min incubation time + 2h incubation time - Lyophilized conjugate New features have been introduce in the bioelisa HIV 1+2 Ag/Ab, compared to the existing Biokit assay: Two new monoclonal antibodies and a new gp41 protein for HIV-1 detection. The new bioelisa HIV 1+2 Ag/Ab improves the performance of previous Biokit assay in terms of analytical sensitivity for p24 detection. Limit of detection is superior to the most important competitors: Murex-Diasorin and Biorad. Specificity has been also improved by means of the new gp41. The large evaluation with blood donor reported a level of specificity not overcome neither Murex-Diasorin nor Biorad. The two kit presentations: 96 and 480 Test adjust better to the customer needs As per the Biokit internal and external results, the bioelisa 1+2 Ag/Ab has to be considered as a new state-of-the-art assay for HIV detection. Page 11 of 15

Launch package: CE Mark Package insert External Evaluations Paul Erhlich Institute Germany Central tropical diseases Establiment Français du Sang (EFS) Page 12 of 15

CE MARK Page 11 of 13

PACKAGE INSERT Page 12 of 13

bioelisa READ HIGHLIGHTED CHANGES bioelisa HIV-1+2 Ag/Ab 3000-1172 1 x 96 TESTS 3000-1173 5 x 96 TESTS (480) ELISA test for the detection of antibodies to Human Immunodeficiency Viruses (HIV) type 1 (group M-O) or type 2 and HIV-p24 antigen in human serum or plasma samples in clinical laboratories and as a first-line screening assay in blood centers. Summary Serological evidence of infection with HIV may be obtained by testing for presence of HIV antigens or antibodies in serum or plasma of individuals suspected for HIV infection. Antigen can generally be detected during both acute phase and the symptomatic phase of AIDS. The antibodies to HIV-1 and/or HIV-2 can be detected throughout virtually the whole infection period, starting at or shortly after the acute phase and lasting till the end stage of AIDS. HIV fourth generation ELISA allows the simultaneous determination of HIV p24 antigens and HIV-1/HIV-2 antibodies reducing therefore the diagnostic window and allowing an early diagnosis of acute HIV infection. Apart from sexual transmission, the principal route of infection with HIV is blood transfusion. HIV can be present in both cellular and cell-free fractions of human blood. Therefore, all donations of blood or plasma should be tested due to the risk of HIV transmission through contaminated blood. This can be effectively achieved by testing for antibodies to HIV and p24 antigen by using a highly sensitive ELISA test. Principle bioelisa HIV-1+2 Ag/Ab is a fourth generation enzyme immunoassay for the simultaneous qualitative detection of HIV p24 antigen and antibodies to HIV-1 (groups M and O) or HIV-2. Antigens representing epitopes of HIV-1 gp41 and HIV-2 gp36 are coated onto microplate wells together with monoclonal antibodies against HIV p24. In the first step of the immunoassay, serum or plasma samples are added to these wells along with the sample diluent containing biotinylated anti-p24 antibodies. If specific HIV-1 and/or HIV-2 antibodies are present in the sample, they will form stable complexes with the HIV antigens attached to the wells. If p24 antigen is present in the sample, it will bind simultaneously to the antibodies in the well and to the biotinylated antibodies present in the sample diluent. The microwells are then washed to remove unbound serum proteins. A set of biotinylated antigens are added to the wells, these antigens will bind to the specific captured antibodies. After a second wash to remove unbound conjugate, a second conjugate of Streptavidin- immunocomplex, the colourless Chromogen is oxidised by the bound conjugate to a blue coloured product. The blue colour changes to yellow after stopping the reaction with sulphuric acid. The intensity of colour can be measured and is proportional to the concentration of anti-hiv 1/2 antibodies or HIV p24 antigen present in the sample. Wells containing negative samples remain colourless. Components 1. MCPL MICROPLATE: 12 x 8 wells coated with antigens representing epitopes of HIV-1 gp41 and HIV-2 gp36 and monoclonal antibodies against p24. Plates are sealed into aluminum pouch with desiccant. 2. CONTROL + 1 HIV-1 POSITIVE CONTROL: Diluted human serum containing antibodies to HIV-1 (Heat inactivated). Contains 0.1% ProClin TM preservative. Ready to use. 3. CONTROL + 2 HIV-2 POSITIVE CONTROL: Diluted human serum containing antibodies to HIV-2 (Heat inactivated). Contains 0.1% ProClin TM preservative. Ready to use. 4. CONTROL + Ag HIV POSITIVE CONTROL p24: Diluted human serum spiked with recombinant HIV-1 p24 antigen. Contains 0.1% ProClin TM preservative. Ready to use 300 as 300 as 300 as 5. CONTROL NEGATIVE CONTROL: Diluted human serum negative for antibodies to HIV-1/2 and HIV p24 antigen. Contains 0.1% ProClin TM 300 as preservative. Ready to use. 6. DIL SAMP SAMPLE DILUENT: Biotinylated monoclonal antibodies against HIV p24 diluted in Tris-buffer. Contains 0.05% ProClin TM 300 as preservative and red dye. Ready to use. BIOKIT, S.A. - Can Malé s/n - - Barcelona - SPAIN 1 0843 3000-1172_R00_05.2014_eng.docx

bioelisa 7. CONJ 1 CONJUGATE READY TO USE: Phosphate buffer containing HIV-1 and HIV-2 biotinylated antigens. Contains 0.05% ProClin TM preservative and blue dye. Ready to use. 300 as 8. DIL CONJ 2 CONJUGATE DILUENT 2: Phosphate buffer containing protein stabilizers, 0.05% ProClin TM 300 as preservative and yellow dye. 9. CONJ 2 101X CONCENTRATE CONJUGATE 2 (101X): Contains peroxidase labelled streptavidin. To be diluted 1:101 in conjugate diluent 2 before use. 10. SUBS TMB SUBSTRATE-TMB: Contains 3,3', 5,5'-Tetramethylbenzidine (TMB) and hydrogen peroxide. Ready to use. 11. WASH SOLN 25x CONCENTRATE WASHING SOLUTION: Concentrate PBS buffer ph 7.4 (25x). Contains Tween-20 as detergent, <0.2% ProClin TM 300 as preservative To be diluted 1:25 in distilled or deionised water before use. 12. H 2 SO 4 0.8N STOPPING SOLUTION: 0.8N sulphuric acid. Ready to use. 13. SEALS ADHESIVE SEALS: To cover the microplate during incubations. 14. BAG RESEALABLE BAG: For storage of unused strips. Precautions bioelisa HIV-1+2 Ag/Ab is intended for IN VITRO diagnostic use. For professional use only. WARNING: The Negative Control, HIV-1 Positive Control, HIV-2 Positive Control, HIV p24 Positive Control, Sample Diluent, Conjugate 1, Conjugate Diluent 2 and Concentrate Washing Solution contain <0.2% ProClin TM 300 as preservative. Hazard statements H317: May cause an allergic skin reaction. Precautionary statements P280: Wear protective gloves/protective clothing/eye protection/face protection. P302+P352: IF ON SKIN: Wash with plenty of soap and water. P333+P313: If skin irritation or rash occurs: Get medical advice/attention. P363: Wash contaminated clothing before reuse. P501: Dispose of contents/container in accordance with local/regional/national/international regulations. WARNING: POTENTIALLY BIOHAZARDOUS MATERIAL. All human source material used in the preparation of this product was found to be negative for the presence of HCV antibodies, Treponema pallidum antibodies, hepatitis B surface antigen and HIV 1/2 antibodies (except for the Positive Controls 1 and 2), using a commercial licensed method. Nevertheless, because no test method can offer complete assurance of the absence of infectious agents, this product should be handled with caution: - Avoid contact of reagents with the eyes and skin. If that occurs, wash thoroughly with water. - Wear gloves. - Do not pipette by mouth. - Do not smoke. - Dispose all used materials in a suitable biohazardous waste container. Remains of samples, controls, aspirated reagents and pipette tips should be collected in a container for this purpose and autoclaved 1-hour at 121 C or treated with 10% sodium hypochlorite (final concentration) for 30 min before disposal. (Remains containing acid must be neutralised prior to the addition of sodium hypochlorite). BIOKIT, S.A. - Can Malé s/n - - Barcelona - SPAIN 2 0843 3000-1172_R00_05.2014_eng.docx

bioelisa Handling instructions: - Adjust washer to the plate used (flat bottom) in order to wash properly. - Do not mix reagents from different lots. - Do not use reagents after the expiration date. - Do not use the reagent if you observed any change in appearance of components included in the kit. - Extreme care should be taken to avoid microbial contamination and cross contamination of reagents. - Use a new pipette tip for each specimen and each reagent. - Do not exchange reagents from different lots or use reagents from other commercially available kits. The components of the kit are precisely matched for optimal performance of the tests. - CAUTION - CRITICAL STEP: Allow the reagents and specimens to reach room temperature (20-25 C) before use. Shake reagent gently before use. Return at 2-8 C immediately after use. - The enzymatic activity of conjugate 2 might be affected by dust and reactive chemical substances like sodium hypochlorite, acids, alkalis etc. Do not perform the assay in the presence of these substances. - It is very important to keep the Substrate-TMB solution in a well-sealed container and avoid light exposure. - Soaps and/or oxidising agents remaining in containers used for the Substrate-TMB solution can interfere with the reaction. If glass containers or re-used plastic containers are used, they should be washed with 1N sulphuric or hydrochloric acid, rinsed well with distilled water and dried before use. We recommend using disposable plastic containers. Storage and stability The components will remain stable through the expiration date shown on the label if stored at 2-8 C. The bag containing the microplate should be brought to room temperature before opening to avoid condensation in the wells. Once opened the bag, the microplate wells are stable for 3 months at 2-8 C in the plastic bag tightly sealed, along with the silicagel. Discard any microplate sealed without the silicagel. Once diluted, the washing solution is stable for 6 days only if stored at 2-8 C. Do not use diluted washing solutions if stored overnight at room temperature. Once diluted, the conjugate 2 is stable for 30 days at 2-8 C. Store the Substrate-TMB solution in the dark. Available packaging - 1 plate kit (1x96 tests), REF 3000-1172. Contains: 1 plate, 1 x 2 ml negative control, 1 x 2 ml HIV-1 positive control, 1 x 2 ml HIV-2 positive control, 1 x 2 ml HIV-1 p24 positive control, 1 x 14 ml sample diluent, 1 x 25 ml conjugate 1,1 x 0.250 ml conjugate 2 (101x), 1 x 25 ml conjugate 2 diluent, 1 x 20 ml substrate-tmb, 1 x 12 ml stopping solution, 2 x 50 ml washing solution (25x), 1 plastic bag and adhesive seals. - 5 plates kit (5 x 96 tests), REF 3000-1173. Contains: 5 plates, 1 x 4 ml negative control, 1 x 4 ml HIV-1 positive control, 1 x 4 ml HIV-2 positive control, 1 x 4 ml HIV-1 p24 positive control, 1 x 70 ml sample diluent, 1 x 120 ml conjugate 1,1 x 1.2 ml conjugate 2 (101x), 1 x 120 ml conjugate 2 diluent, 1 x 100 ml substrate-tmb, 1 x 60 ml stopping solution, 3 x 100 ml washing solution (25x), 1 plastic bag and adhesive seals. Material required not provided - Distilled or deionised water. - Disposable gloves and timer. - Appropriate waste containers for potentially contaminated materials. - Disposable V-shaped troughs. - Dispensing system and/or pipette (single or multichannel) and disposable pipette tips. - Absorbent tissue or clean towel. - Dry incubator or water bath, 37 ± 1 C. - Microplate reader with a 450 nm filter. Reference filter of 620 nm or 630 is advisable. - Manual or automated wash system. Sample collection Use fresh serum with or without Serum separator tube (SST), EDTA plasma, Acid-citrate-dextrose (ACD) plasma, Lithium heparin plasma, Sodium heparin plasma, Sodium citrate plasma, Potassium oxalate/sodium fluoride plasma, CPD and CPDA plasma. Other anticoagulants should be evaluated before use. Samples can be stored at 2-8 C for 8 days. For longer periods, samples should be frozen (-20 C). Avoid repeated freezing and thawing (maximum 4 freeze-thaw cycles). Specimens showing visible particulate matter should be clarified by centrifugation. Serum or plasma samples should not be heat inactivated, since that may cause incorrect results. BIOKIT, S.A. - Can Malé s/n - - Barcelona - SPAIN 3 0843 3000-1172_R00_05.2014_eng.docx

bioelisa Automatic processing Automated or semi-automated assay may be used with different instruments. It is very important to validate any automated system to demonstrate that the results obtained for samples are equivalent to the ones obtained using the manual assay. It is recommended that the users validate periodically the instrument. If there is any difficulty in the programming and setting of Biokit automatic processors, please contact your distributor. PROCEDURE Previous operations Allow all the reagents to reach room temperature (20-25 C) before running the assay. Gently mix all liquid reagents before use. Check the washing solution concentrate for the presence of salt crystals. If crystals have formed, resolubilize by warming at 37 C until crystals dissolve. Use distilled or deionized water. Negative and Positive controls should be treated as the samples. Dilute the washing solution 1:25 in distilled water. If the entire plate is used, add 40 ml of concentrate washing solution (25x) to 960 ml of deionised water. If the entire plate is not used, prepare the proportional volume of solution. Dilute the concentrated conjugate 2 1:101 with the conjugate diluent 2 according to table 1. For the 1 plate packaging, if the entire plate is to be used, add 250 μl of concentrate conjugate directly to the bottle containing 25 ml of conjugate diluent. Mix gently. TABLE 1 Strips required 1 2 4 6 8 10 12 Conjugate diluent 2 ml 2.0 4.0 8.0 12.0 16.0 20.0 25.0 Concentrate conjugate 2 μl 20 40 80 120 160 200 250 Assay procedure 1. Use only the number of strips required for the test. Reserve 7 wells for blank and controls. One well as Blank (e.g. A1), three wells as negative control (e.g. B1, C1, D1) and, at least, one well of each of the positive controls: positive control HIV-1 (e.g. E1), positive control HIV-2 (e.g. F1) and positive control HIV p24 (e.g. G1). 2. Add 100μL of sample diluent to all the wells except the Blank. 3. Add first 100 l of samples, and last 100 l of negative and each of the positive controls into their respective wells. 4. Cover the plate with an adhesive seal, mix gently and incubate at 37 C ± 1 C for 60 minutes with a tolerance range of 58 65 minutes 5. Remove and discard the adhesive seal. Aspirate the contents of the wells and fill them completely (approximately 350 μl) with the diluted washing solution. Repeat the process of aspiration and washing 4 more times, 5 (five) cycles in total. Ensure that each column of wells soaks for at least 15 seconds before the next aspiration cycle. 6. Transfer 200 μl of conjugate 1 into each well, except the one reserved for the substrate blank. Avoid bubbles upon addition. 7. Cover the microplate with an adhesive seal and incubate at 37 C ± 1 C for 30 minutes with a tolerance range of 28 35 minutes. 8. Remove and discard the adhesive seal. Aspirate and wash the wells as in the previous washing step for 3 (three) wash cycles. 9. Transfer 200 μl of working conjugate 2 into each well, except the one reserved for the substrate blank. Avoid bubbles upon addition. 10. Cover the microplate with an adhesive seal and incubate at 37 C ± 1 C for 30 minutes with a tolerance range of 28 35 minutes. 11. Remove and discard the adhesive seal. Aspirate and wash the wells as in step 5, for 5 (five) wash cycles. BIOKIT, S.A. - Can Malé s/n - - Barcelona - SPAIN 4 0843 3000-1172_R00_05.2014_eng.docx

bioelisa 12. Add 150 μl of Substrate-TMB solution to each well, including the blank. 13. Incubate at room temperature (20-25 C) for 30 minutes with a tolerance range of 28 35 minutes, protected from direct light exposure 14. Stop the reaction by adding 100 μl of stopping solution in the same sequence and time intervals as for the Substrate-TMB. 15. Blank the reader at 450 nm with the blank well and read the absorbance of each well within 30 minutes. It is recommended to read in bichromatic mode using a 620-630 nm reference filter. Quality control Results of an assay are valid if the following criteria are accomplished: 1. The Absorbance value of the blank well must be 0.100. 2. The Absorbance value of the 0.900 after subtracting the blank. If not the run should be repeated. 3. Each individual absorbance value of the negative control must 0.120 after subtracting the blank. If one of the negative control values does not meet the Quality Control criteria, it should be discarded and the mean value calculated again using the remaining two values. If two or more negative control Absorbance values do not meet the Quality Control Range specifications, the test is invalid and must be repeated. Results 1. Subtract the blank from each of the valid negative controls. Calculate the mean absorbance of the negative control (NCx). Calculate the cut-off value by adding 0.170 to the mean absorbance of the negative control. Cut-off = NCx + 0.170 Example: NCx = 0.011 cut-off value = 0.011 + 0.170 = 0.181 2. Divide the absorbance of the sample by the cut-off value. Positive: ratio absorbance/cut-off 1.0 Negative: ratio absorbance/cut-off 0.9 Equivocal: ratio absorbance/cut-off 0.9 < 1.0 Interpretation of results Negative Results: Specimens with an absorbance to cut-off ratio lower than 0.9 are considered non reactive which indicates that no anti-hiv 1 and/or HIV-2 antibodies or HIV p24 antigen have been detected with bioelisa HIV-1+2 Ag/Ab kit. Positive Results: Specimens giving an absorbance equal to or greater than the cut-off value are considered initially reactive, which indicates that anti-hiv 1/2 antibodies or HIV p24 antigen have probably been detected using bioelisa HIV-1+2 Ag/Ab. All initially reactive specimens should be retested in duplicate to confirm the initial result. Equivocal: Specimens with an absorbance to cut-off ratio between 0.9 and 1.0 are considered equivocal and retesting of these specimens in duplicate is required to confirm the initial results. - If, after retesting the initially reactive samples, both wells report negative results (S/CO < 0.9), these samples should be considered as non-repeatable positive (or, false positive) and recorded as negative. - If, after retesting in duplicate, one or both wells report positive results, the final result from this ELISA test should be recorded as repeatedly reactive. Repeatedly reactive specimens can be considered positive for antibodies to HIV 1/2 or p24 antigen, therefore the patient is probably infected with HIV1 or HIV2. - After retesting in duplicate, samples with values close to the cut-off value should be interpreted with caution and considered as equivocal, or uninterpretable for the time of testing. - Follow-up, confirmation and supplementary testing of any positive specimen with other analytical system (e.g. WB, PCR) is required. Clinical diagnosis should not be established based on a single test result. It should integrate clinical and other laboratory data and findings. BIOKIT, S.A. - Can Malé s/n - - Barcelona - SPAIN 5 0843 3000-1172_R00_05.2014_eng.docx

bioelisa Limitations of the procedure - Optimal assay performance requires strict adherence to the assay procedure described. Deviation from the procedure may lead to aberrant results. - As in all sensitive immunoassays, there is the possibility that non-repeatable positive results occur. - Positive results must be confirmed with another available method and interpreted in conjunction with the patient clinical information. - Negative results obtained with bioelisa HIV-1+2 Ag/Ab are considered negative for antibodies to HIV-1/2 and p24 antigen and further testing is not required. - - This assay cannot be used to test pooled (mixed) plasma. bioelisa HIV-1+2 Ag/Ab has been evaluated only with individual serum or plasma specimens. - bioelisa HIV-1+2 Ag/Ab is a qualitative assay and the results cannot be used to measure concentrations of antibodies to HIV or p24 antigen. This assay cannot distinguish between infections with HIV-1 and HIV-2. - A negative result does not exclude the possibility of exposure or infection with HIV. Expected results The number of positive results depends on the disease incidence in the geographic area and the type of tested population. In the world, the HIV incidence in people older than 15 years varies from 0.1% in Australia, New Zeeland and Asia Pacific, 0.3% in Western Europe, North Africa and Middle East, 0.5% in East Europe, Central Asia and Latin America, 0.6% in North America and South Asia and 9% in sub-saharan Africa. In the same country, incidence also varies enormously according to the tested population. In Western Europe, HIV antibodies prevalence in blood bank donations varies from 0 to 5 cases over 100,000 donations, while the incidence among prisoners, sex workers and drug abusers may easily reach 20% in these risk populations. Performance characteristics Analytical Sensitivity The limit of detection has been estimated below 0.5 IU/ml by testing the WHO HIV p24 Antigen, First International Reference NIBSC code 90/636 with two different lots during the internal evaluation. Additionally, the limit of detection has been also estimated below 12.5 pg/ml by testing BIORAD HIV-1 Antigen Standard 72217 with two different lots during the internal evaluation. Sensitivity - A total of 537 HIV-1 positive samples were evaluated with bioelisa HIV 1+2 Ag/Ab, among them: 28 HIV-1 fresh-same-day collected samples, 85 HIV-1 and 4 HIV-1 Subtype O samples in an European reference lab; 380 HIV -1 samples were evaluated at Biokit and 40 HIV-1 non B subtype samples (and at least 3 samples of each HIV-1 subtype) were evaluated in another European reference lab. bioelisa HIV 1+2 Ag/Ab detected all 537 samples. The sensitivity found was100% (537/537). - A total of 115 HIV-2 positive samples were evaluated with bioelisa HIV 1+2 Ag/Ab, among them: 27 HIV-2 samples in an European reference lab and 88 HIV-2 samples were evaluated at Biokit. bioelisa HIV 1+2 Ag/Ab detected all 115 samples. The sensitivity found was 100% (115/115). - A total of 61 HIV-1 p24 Ag positive samples were evaluated also with bioelisa HIV 1+2 Ag/Ab, among them: 15 samples in an European reference lab, 16 samples were evaluated in another European reference lab and 30 samples at Biokit. bioelisa HIV 1+2 Ag/Ab detected all 61 samples. The sensitivity found was 100% (61/61). - Additional sensitivity studies were carried out by testing a total of 31 seroconversion panels at Biokit and a European reference lab. Results obtained were comparable to the most sensitive commercial method for HIV 1+2 antigen and antibody detection. - 16 HIV Seroconversion Panels from BBI/Seracare and Zeptometrix were evaluated at Biokit. bioelisa HIV-1+2 Ag/Ab identified correctly all the positive specimens. BIOKIT, S.A. - Can Malé s/n - - Barcelona - SPAIN 6 0843 3000-1172_R00_05.2014_eng.docx

bioelisa bioelisa HIV Ag/Ab HIV p24 HIV Ab Assay Seroconversion Panel HIV Ag/Ab Assay antigen NAT First sample detected positive in the panel BB1-PRB965-n=6 2 2 4 2 1 ZEPT-12007-n=9 4 4 5 ND 4 BBI-PRB954-n=7 6 6 7 6 4 BBI-PRB932-n=9 4 ND 4 4 4 ZEPT-9017-n=11 1 4 3 1 1 ZEPT-9014-n=7 1 1 3 1 1 ZEPT-9012-n=8 5 6 7 5 1 ZEPT-9032-n=14 8 8 7 8 1 ZEPT-6243-n=10 7 7 8 7 1 ZEPT-9022-n=9 7 8 9 7 1 ZEPT-9018-n=11 9 9 10 9 1 BBI-PRB939-n=9 6 ND 8 6 5 BBI-PRB956-n=5 4 4 5 4 2 BBI-PRB945-n=6 3 4 4 3 1 BBI-PRB955-n=4 2 2 4 2 1 BBI-PRB958-n=6 3 3 5 3 1 - In the previous table it can be observed the first sample detected as positive in each panel evaluated with bioelisa HIV-1+2 Ag/Ab in comparison with 4 th Generation assays (Ag/Ab Assays), 3 rd Generation assays (Ab Assays), assays for HIV p24 antigen and HIV RNA NAT assays. bioelisa HIV-1+2 Ag/Ab showed excellent sensitivity to detect early antigen or antibodies in 16 HIV Seroconversion Panels evaluated. - Another study was conducted to assess the sensitivity in 32 early seroconversion samples. These type of samples are characterized to be positive to the presence of HIVp24 Ag and/or positive by NAT (Nucleic Acid Testing), not detected by all HIV 3rd generation methods and to be negative or Indeterminate by Western-Blot methods. bioelisa HIV 1+2 Ag/Ab detected all 32 samples. The sensitivity found was 100% (32/32). - Finally, 50 cell culture supernatans from HIV cell cultures, including different HIV-1 subtypes and HIV-2 were evaluated in a European Reference Lab. bioelisa HIV 1+2 Ag/Ab detected all 50 samples. The sensitivity found was100% (50/50). Specificity The specificity was evaluated by testing a total of 6465 unselected blood donors' samples at three different sites. - In the first site 3045 unselected samples including 684 first donors were tested. From this total, 4 samples were reactive. The specificity obtained in this study was 99.87% (3041/3045). - In the second site 500 unselected samples from Blood Bank were tested. All 500 samples were negative. The specificity was 100% (500/500) - An internal evaluation of 2920 unselected samples from Blood Bank was performed. From this evaluation, 6 samples were reactive. The specificity obtained in this study was 99.79%. (2914/2920) A detailed summary of these specificity studies is shown in the table below. BIOKIT, S.A. - Can Malé s/n - - Barcelona - SPAIN 7 0843 3000-1172_R00_05.2014_eng.docx

bioelisa Evaluation 1 (3045 sera) Initial Reactive Repeated Reactive False Positive Specificity % Serum 6 4 4 99.87 Evaluation 2 (500 sera) Initial Reactive Repeated Reactive False Positive Specificity % Serum 0 0 0 100 Internal Evaluation (1000 sera + 1920 plasmas) Initial Reactive Repeated Reactive False Positive Specificity % Plasma 8 6 6 99.69 Serum 1 0 0 100 Total 9 6 6 99.79 - In addition 200 samples from hospitalized patients have been tested and compared to a reference test. 194 samples were found negative by both tests and six samples were reactive. The six samples were also reactive with other ELISA tests. A specificity of 100% was obtained in this study (194/194). Precision Intra-assay reproducibility: The coefficient of variation obtained for the absorbance values of a HIV-1 positive sample assayed in a minimum of 40 replicates was 3.42%, 5.79% and 4.71% in three lots studied. The coefficient of variation obtained for the absorbance values of a HIV-2 positive sample assayed in a minimum of 40 replicates was 6.39%, 5.91% and 8.38% in three lots studied. The coefficient of variation obtained for the absorbance values of a HIV-1 p24 positive sample assayed in a minimum of 40 replicates was 4.08%, 3.44% and 3.71% in three lots studied. Inter-assay reproducibility: Four positive samples of different levels were tested in 15 different assays. The coefficients of variation obtained for the ratios absorbance/cut-off of the four samples were 4.87%, 5.37%, 7.49% and 7.54%. Interferences Interference by addition No interference has been found for haemoglobin (500 mg/dl), bilirubin (20 mg/dl), human Albumin (5 g/dl) and triglycerides (500 mg/dl). Cross-reactivity To evaluate possible interferences, 125 potential cross-reacting specimens were analyzed. Among those samples, 4 samples (RF+), 4 samples positive for anti-nuclear antibodies (ANA), 5 samples Mononucleosis, 89 samples from other related infectious diseases and 23 samples from pregnant women. No evidence of cross reactivity was found in the samples evaluated. The 2 anti-toxoplasma IgM samples that tested positive were also positive by other HIV assays. Potential interfering samples = 125 RF (Rheumatoid factor) - 4 Elevated IgG - 4 ANA (anti-nuclear Antibodies) - 4 Elevated IgM - 5 Mononucleosis - 5 HSV 1 IgG (Herpes Simplex Virus) - 5 Pregnant women- 23 (including 7 multiparous) HSV 2 IgG (Herpes Simplex Virus) - 4 Syphilis Ab - 6 anti-cmv IgG (Cytomegalovirus) - 5 anti-ebv (Epstein-Barr Virus) - 3 anti-cmv IgM (Cytomegalovirus) - 7 anti-rubella (Rubella Virus) - 4 anti-htlv (Human T-lymphotropic Virus) - 4 anti-vzv IgG (Varicela Zoster Virus) - 2 anti-hcv (Hepatitis C Virus) - 8 anti-vzv IgM (Varicela Zoster Virus) - 2 anti-e.coli (Escherichia coli)-5 anti-toxoplasma IgG (T. gondii) - 10 HBsAg (Hepatitis B Antigen) - 8 anti-toxoplasma IgM (T. gondii)-7 BIOKIT, S.A. - Can Malé s/n - - Barcelona - SPAIN 8 0843 3000-1172_R00_05.2014_eng.docx

bioelisa bioelisa: Troubleshooting guide Problem Possible causes Solution 1. Controls out of validation. 2. No colour or only a light colour developed at the end of the assay. 1a. Incorrect temperature, timing or pipetting. 1b. Improper preparation of reagents, error of dilution, reagents not well mixed. 1c. Cross-contamination of controls. Check procedure. Repeat assay. Check procedure. Repeat assay. Pipette carefully. Do not interchange caps. Repeat assay. 1d. Incorrect reading filter. Check that the wavelenght of the filter used is 450 nm. If no reference 620-630 nm is used, absorbance increases approximately 0.050. 1e. Interference in the optical pathway. Check the reader. Clean or dry the bottom of microplate. Check for air bubbles. Repeat reading. 1f. Used components from different lots. Do not use components from different lots as they are adjusted for each batch released. 1g. Expired reagents. Check the kit expiry date. Use 2a. One or more reagents not added or added in wrong sequence. 2b. Inactive conjugate. Wrong dilution of concentrate conjugate 2. Improper conservation. 2c. Inactive microplate: Improper conservation. 2d. Inactive Sustrate-TMB: Improper conservation. The container used affects Substrate-TMB stability, cross-contamination with the stopping solution. a non- expired kit. Check procedure. Repeat assay. Check for contamination. Check procedure. Repeat assay. Always keep unused strips in the bag very well closed, with the desiccant inside. Repeat assay. Use disposable containers or wash with acid or ethanol and rinse with deionised water before re-use. Check procedure. Repeat assay. 2e. Too cold reagents. Let reagents reach room temperature before use. BIOKIT, S.A. - Can Malé s/n - - Barcelona - SPAIN 9 0843 3000-1172_R00_05.2014_eng.docx

bioelisa bioelisa: Troubleshooting guide Problem Possible causes Solution 3. Too much colour in all microplate wells. 4. Poor reproducibility or high number of non-repeatable reactive samples. 3a. Contaminated or oxidised Substrate-TMB solution. 3b. Contaminated or improperly prepared reagents. 3c. Contaminated washing solution (1x). 3d. Insufficient washing or washing not consistent: filling volume and/or aspiration insufficient or not uniform. Insufficient number of washing cycles, contaminated device. 3e. Using of a washing solution from other manufacturer. Check that substrate is colourless, discard if blue. Use acid or ethanol washed or disposable containers. Repeat assay. Check for contamination: turbid aspect. Check dilutions. Repeat assay. Check the quality of distilled or deionised water used for dilution. Repeat assay. Check the washing device. Fill wells with washing solution close to the top, aspirate completely. Increase the number of wash cycles. Use only the washing solution supplied with the kit. 4a. Washing problems. See 3c, 3d, 3e. 4b. Uncalibrated pipettes or tips not well fitted. Improper pipetting. 4c. Reagents too cold or not well mixed before using. 4d. Air currents over the microplate during incubations. 4e. Too long time for addition of samples and/or reagents. Inconsistency in time intervals. Air bubbles. 4f. Interference in the optical pathway. Use only calibrated pipettes, with well fitted tips and pipette carefully, without bubbles and splashing. Repeat assay. Equilibrate reagents to room temperature and mix thoroughly before using. Keep the microplate protected from air currents. Develop consistent and uniform technique. See 1e. BIOKIT, S.A. - Can Malé s/n - - Barcelona - SPAIN 10 0843 3000-1172_R00_05.2014_eng.docx

EXTERNAL EVALUATION Page 13 of 13

Prüflabor für In-vitro Diagnostika beim Paul-Ehrlich-Institut (PEI-IVD) Paul-Ehrlich-Straße 51-59 D-63225 Langen Evaluation Report for bioelisa HIV-1+2 AG/AB Manufacturer: Biokit S.A. Can Malé s/n, 08186 Lliçà d Amunt, Barcelona Spain Dr. Sigrid Nick Prüflabor für In-vitro-Diagnostika Paul-Ehrlich-Institut Phone: +49 6103 777008 Fax: +49 6103 771234 E-mail: Sigrid.Nick@pei.de

bioelisa HIV-1+2 AG/AB Purpose of the Evaluation: The purpose of the evaluation of the bioelisa HIV-1+2 AG/AB carried out by the Prüflabor for In-vitro Diagnostic Medical Devices at the Paul-Ehrlich-Institut (PEI-IVD) was to assess the clinical sensitivity of the device using 15 commercial HIV-1 seroconversion panels, 100 HIV-1 positive samples, including 25 fresh samples (<24 h of collection), a HIV-1 p24 antigen positive subtype panel (14 samples of HIV-1 subtypes A, B, C, D, E, F, G, H and O), 4 HIV-1 O samples and 25 HIV-2 samples. In addition, the analytical sensitivity was to be determined using the HIV-1 p24 reference material of the Paul-Ehrlich-Institut (PEI). The specificity was to be evaluated using 500 German blood donor sera to contribute to the performance requirements as specified in Table 1 of the Common Technical Specifications (CTS). Materials and Methods: Description of the Device The bioelisa HIV-1+2 AG/AB is a non-automated enzyme-linked immunosorbent assay (ELISA) for the simultaneous detection of HIV-1 p24 antigen and IgM, IgA as well as IgG antibodies to HIV-1 and HIV-2 in human serum and plasma. The instructions for use specify that the assay is intended for the detection of HIV infection by clinical laboratories and for screening in blood donation centres. According to the labels and to the instructions for use, the kits contain the following components and show the following features: Table 1. Features of the bioelisa HIV-1+2 AG/AB Test Product name: Id.-no.: Manufacturer: bioelisa HIV-1+2 AG/AB 3000-1172 (96 tests), 3000-1173 (480 tests) Biokit S.A. Can Malé s/n, 08186 Lliçà d Amunt, Barcelona, Spain Instructions for Use (version): V.2 Intended use: Detection of HIV-1 p24 and HIV-1- and HIV-2-specific antibodies in human serum or plasma samples; blood screening assay Sample volume: 100 µl Number of determinations: 96 or 480 tests per kit Lot no.: J-1912 Expiry date: 08.06.2013 Storage temperature: 2-8 C Assay format: ELISA Kit components: - Breakable microtitre plates coated with anti-hiv-1 p24 antigen monoclonal mouse antibodies and recombinant HIV-1 (gp41) and HIV-2 (gp36) antigens PEI-IVD Page 2 of 34

bioelisa HIV-1+2 AG/AB Measuring/ reference wave lengths: Control specifications: Blank HIV-1 positive control - Blank HIV-2 positive control - Blank HIV p24 positive control - Blank NC - Blank - CONTROL + 1: HIV-1 positive control (PC) - CONTROL + 2: HIV-2 positive control - CONTROL + Ag: HIV positive control p24 (recombinant purified HIV-1 p24 antigen diluted in human serum) - CONTROL - : Negative control (NC) - Sample diluent - CONJ 1 (biotinylated HIV-1 and HIV-2 antigens) - DIL CONJ 2: conjugate diluent 2 - CONJ 2 (101x concentrate; Peroxidase labelled streptavidin) - TMB substrate - Washing solution concentrate (25x) - Stopping solution - Adhesive seals - Re-sealable bag - Instructions for use 450 / 620-630 nm Mean Absorbances (A): 0,050 0,500 0,500 0,500 Cut-off (CO): A NCmean + 0,170 0,100 (each individual value) Assay Performance Test kits were provided by Biokit, Spain. Testing was performed from December 2012 through January 2013. At the time of the investigation, the device was not yet CE-marked. Testing procedure of the bioelisa HIV-1+2 AG/AB strictly followed the instructions for use (IFU). In brief, test components and samples were allowed to reach room temperature. Washing solution and conjugate solution no. 2 were prepared in the required volumes. Sample diluent was pipetted to the wells except for the Blank well. Then 100 µl of the Controls or of the test samples were added to the wells of the microtitre plate and mixed. Thereafter, the plates were sealed and transferred to the incubator without delay. After incubation at 37 C for 1 h, plates were washed five times with 400 µl washing solution in the overflow modus using a HydroFlex washer (Tecan). Soak time was 30 sec. After the addition of 200 µl of the conjugate solution no. 1, plates were incubated for 30 min and washed again three times. Then 200 µl of the conjugate solution no. 2 were added to the plates and after sealing plates were incubated for another 30 min and thereafter washed again five times PEI-IVD Page 3 of 34

bioelisa HIV-1+2 AG/AB For colour development 150 µl of the TMB substrate were added, plates were incubated at room temperature as required and then the reaction was stopped with 100 µl sulphuric acid. Absorbance values (A) were read at dual wave lengths 450/620 nm using a Tecan sunrise photometer. All pipetting steps were performed manually. Anti-HIV-1 and anti-hiv-2 positive samples as well as the seroconversion panels were tested in single determination. In case of results that were discrepant from the known sample status or of unusual results of the test kit controls or the samples, the determination was repeated. Determination of Analytical Sensitivity The analytical sensitivity was determined using the PEI HIV-1 p24 reference material. The concentration range is indicated in Table 2 below. Each dilution step was tested in double determination and tested three times apart. Table 2. HIV-1 p24 Antigen Standard Preparations Reference Material Description Concentration Range PEI HIV-1 p24 reference material Recombinant HIV-1 p24 expressed in E. coli and provided by Roche Diagnostics, Germany 100 pg/ml 1,60 pg/ml Anti-HIV-1 Positive Samples Eighty-five Anti-HIV-1 positive samples were obtained from the HIV Center of the University hospital of Frankfurt, Germany, in September 2008. Samples were tested for HIV at the time of receipt. Comparative results were available from the Murex HIV Ag/Ab Combination (now DiaSorin), the ABBOTT PRISM HIV O Plus, the ABBOTT ARCHITECT HIV Ag/Ab or the AxSYM HIV-Ag/Ab Combo test. After pretesting, samples were stored at -20 C until use. In addition, 28 Anti-HIV-1 positive samples were tested at the day of receipt (within 24 h of sample collection). Anti-HIV-1 O Positive Samples To investigate the ability of the bioelisa HIV-1+2 AG/AB to detect HIV-1 O, 4 positive samples were tested. Three samples were collected in Cameroun and were tested neat. One sample (W 92005) originated from Germany. Sample W 92005 was tested in a 1:100 dilution. Anti-HIV-2 Positive Samples In total, 25 anti-hiv-2 positive samples were included in the investigation (Appendix 2). Samples obtained from SeraCare were collected in Guinea Bissau, West Africa. According to the information provided by SeraCare, these samples were positive by the Siemens ADVIA Centaur PEI-IVD Page 4 of 34

bioelisa HIV-1+2 AG/AB HIV Enhanced (EHIV) assay and yielded overflow results (index > 50). The HIV-1/-2 type differentiation was carried out by SeraCare with an Anti-HIV-1/2 rapid test (Genie Multispot HIV- 1/2, Bio-RAD). Final discrimination between HIV-1 and HIV-2 infection was performed at PEI- IVD using the CHIRON RIBA HIV-1/HIV-2 SIA supplemental assay. The latter investigation of PEI classified 21 samples as unequivocally HIV-2 positive, three samples exhibited equal reactivity with HIV-1 and HIV-2 antigens. For these 3 samples it remains unclear whether the results are due to cross-reactivity or to HIV-1 and HIV-2 double infection. Further two anti-hiv-2 positive PEI samples were included. One sample was collected in Germany; the second sample was obtained from SeraCare (former BBI). HIV-1 p24 Subtype Samples To investigate the ability to detect different HIV-1 p24 subtypes, 14 samples of the former Nationales Referenzzentrum (NRZ) für Retroviren, Virologisches Institut - Klinische und Molekulare Virologie of the Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany were tested. At the time of receipt, the NRZ HIV-1 subtype panel included 11 group M isolates (1 x A, 1 x B, 1 x C, 3 x D, 2 x CRF01_AE ( E ), 2 x F, 1 x G, 1 x H) and two group O isolates. Virus isolates were originally obtained via the Programme EVA, Centre for AIDS Reagents, National Institute for Biological Standards and Control (NIBSC), UK. These virus isolates were propagated on cells and supernatants were diluted in HIV-negative plasma to a concentration of about 100,000 HIV-1 RNA copies/ml. The isolates were quantified by both the branched DNA assay (Siemens) and different real time HIV-1 PCR assays (by Abbott or Siemens). Precise concentrations (copies/ml) are given in Table 3 below. Table 3. Characteristics of the HIV-1 p24 subtype panel No. HIV 1 Subtype Laboratory ID Production date Viral Load (NRZ) (copies/ml) 1 A A 26.08.2005 126.600 2 B B 24.04.2008 64.100 3 C C 23.09.2005 72.600 4 D D1 28.10.2005 72.000 5 D D2 28.10.2005 128.400 6 D D3 11.11.2005 73.600 7 E E1 23.09.2005 190.000 8 E E2 10.11.2005 100.200 9 F F1 20.01.2006 140.400 10 F F2 24.07.2009 88.800 11 G G 24.04.2008 207.700 12 H H 15.09.2008 124.300 13 O O1 28.05.2009 624.000 14 O O2 27.10.2008 135.500 PEI-IVD Page 5 of 34

bioelisa HIV-1+2 AG/AB HIV-1 Seroconversion Panels In addition, 15 commercial seroconversion panels were tested (Table 4). Seroconversion panels were pre-selected for short bleeding intervals to allow for a good discrimination between sensitive and less sensitive HIV tests. The seroconversion panels were purchased from either SeraCare Life Sciences (37 Birch Street, Milford, MA 01757, USA) or from Zeptometrix Corporation (872 Main Street Buffalo, NY 14202, USA). Table 4. HIV-1 Seroconversion Panels Used for Testing of the bioelisa HIV-1+2 AG/AB 1 2 3 4 5 6 7 Panel No. PRB 960 PRB 963 PRB 966 PRB 967 PRB 969 PRB 971 PRB 972 Donor No. Panel vendor Bleeds per panel HIV-1 p24 Agpositive panel members HIV Ag/Abpositive bleeds 1 st HIVantibody reactive bleed Results of confirmatory or supplemental tests Maximum no. of bleeds reactive by the most sensitive HIV assay n.a. SeraCare 9 8 to 9 8 to 9 n.a. n.a. 2 n.a. SeraCare 7 6 to 7 6 to 7 7 Western blot (WB) negative n.a. SeraCare 10 8 to 10 8 to 10 9 10 indeterminate 3 n.a. SeraCare 6 4 to 6** 4 to 6 4** 5 indeterminate, 6 positive n.a. SeraCare 10 7 to 9** 8 to 10 8** 8 to 10 positive 3 n.a. SeraCare 4 3 to 4** 3 to 4 4** WB negative 2 n.a. SeraCare 6 5 to 6** 5 to 6 5** RIBA negative 2 8 6240* 60772 ZeptoMetrix 9 6244 61694-74 11* (13) 7 to 12 7 to 13 9 10 indeterminate; 11 to 13 positive Zeptometrix 15 13 to 15 13 to 15 15 all negative 2 10 6247 63602 ZeptoMetrix 10 7 to 10 7 to 10 10** all negative 4 11 6248 63331 ZeptoMetrix 7 6 to 7 6 to 7 7 all negative 2 12 9010 65376 ZeptoMetrix 11 9 to 11 9 to 11 10 11 indeterminate 3 13 9025 67996 ZeptoMetrix 12 11 to 12 11 to 12 12 all negative** 2 14 9030 68582 ZeptoMetrix 16 14 to 16 14 to 16 16 all negative 3 15 9034 66632 ZeptoMetrix 13 11 to 13 11 to 13 13 all negative 3 *Panel 6240: members 1 and 2 could not be tested due to insufficient volume (VNS); ** information taken from the panel vendors sheets; n.a. not applicable; n.d. not determined The results obtained for the bioelisa HIV-1+2 AG/AB were compared to data of current CEmarked HIV-Ag/Ab combination tests with state-of the-art technology (Appendix 6). The HIV- Ag/Ab combination test I exhibits ELISA format. HIV-Ag/Ab combination test II is a chemiluminescence immunoassay; currently, this is the most sensitive test for which comparative data is available at the Testing Laboratory for IVD. Data from RNA detection by NAT (nucleic assay amplification techniques) were taken from the panel vendors sheets which are attached (Attachments 1 and 2). In addition, results from current CE-marked Anti-HIV-1/2 tests as well as confirmatory Western blots or immunoblots were taken into account. PEI-IVD Page 6 of 34 2 3 7

bioelisa HIV-1+2 AG/AB Specificity Specificity was tested using 500 sera from the German Red Cross Blood Donor Service Baden- Wuerttemberg Hessen, Institute for Transfusion Medicine and Immunohaematology Frankfurt/Main. Samples were routinely collected by the Red Cross one day and pretested negative by the Abbott Prism HIV Ag/Ab Combo test. Left-over samples were received that same day and tested by the bioelisa HIV-1+2 AG/AB within three days after receipt. Results Analytical Sensitivity Analytical sensitivity was determined with dilutions of the WHO HIV-1 p24 1 st International reference reagent on three days. Mean analytical sensitivities were 10,45 pg/ml and 9,81 pg/ml and 8,00 pg/ml (Appendix 1) with a grand overall mean of 9,42 pg/ml. Anti-HIV-1 Positive Samples and Anti-HIV-1 O Samples In total 85 samples collected in 2008 and stored at -20 C were tested by the bioelisa HIV-1+2 AG/AB test. A selection criterion for the samples was that results from current CE-marked devices with state-of-the-art performance were already available. All 85 samples were reactive by the bioelisa HIV-1+2 AG/AB (Appendix 2) and showed high S/CO ratios between 15,38 and 25,00. Neither was a false-negative result obtained, nor was any discrepancy from the predetermined sample status observed. In addition, 28 fresh samples that are obtained routinely in the laboratory were tested within 24 h after draw. All fresh samples were clearly reactive and showed S/CO ratios above 16 (Appendix 3). Sensitivity was thus 100% for anti-hiv positive specimens stored frozen or tested fresh. Four Anti-HIV-1 O samples were tested. Samples W 25290, W25291 and W 26633 yielded high S/CO ratios in the overflow range (Table 5). Sample W92005-1 was tested in a 1:100 dilution and was positive, too. Table 5. HIV-1 Subtype O Samples PEI samples Test name bioelisa HIV-1+2 Ag/Ab Manufacturer Biokit S.A. ID-No. 3000-1172 Lot no. J-1912 Test date 29.01.13 Sample No. HIV-1 subtype Origin s/co W 25290 O Cameroun 27,20 W 25291 O Cameroun 27,21 W 26633 O Cameroun 27,21 W92005-1* O Germany 1,52** *diluted 1:100 **mean of three determinations PEI-IVD Page 7 of 34

bioelisa HIV-1+2 AG/AB Anti-HIV-2 Positive Samples Twenty-five anti-hiv-2 positive samples collected in Guinea-Bissau (obtained from SeraCare) were tested. According to the vendor s data the samples are high positive and exhibited overflow reactivity (> 50 index) in the Siemens ADVIA Centaur HIV Enhanced (EHIV). In addition, the samples were confirmed HIV positive by the CHIRON RIBA HIV-1/HIV-2 SIA. Three samples were characterized anti-hiv-1 and anti-hiv-2 positive by the CHIRON RIBA HIV-1/HIV-2; thirty-six samples were classified anti-hiv-2 positive. All 25 samples were detected as reactive by the bioelisa HIV-1+2 AG/AB test and yielded high S/CO ratios (> 17,5) (Appendix 4). Results were thus compliant with the pre-determined sample status. Two further HIV-2 positive samples were tested. Sample 26 is a sample from India (obtained from BioClinical Partners, now ZeptoMetrix) and sample 27 was collected in Germany. Although both additional samples were already pre-diluted, they were also clearly reactive. Overall sensitivity on anti-hiv-2 positive specimens was 100%. HIV-1 p24 Antigen Subtype Samples To investigate the ability of the bioelisa HIV-1+2 AG/AB test to recognise different HIV-1 p24 antigen subtypes, 14 samples from the Institut für Klinische und Molekulare Virologie (National Reference Centre for Retroviruses from April 1996 to September 2012) were investigated. Samples originating from cell culture supernatants were spiked into normal human serum and then tested without further dilution in the bioelisa HIV 1+2 Ag/Ab test. Nine samples were reactive (S/CO ratio 1,0), one sample was in the grey zone (S/CO ratio 0,91) and four samples were negative (S/CO ratio < 0,9). The data is summarized in the table in Appendix 5. Early Seroconversion Sensitivity To assess early seroconversion sensitivity, the bioelisa HIV-1+2 AG/AB test was investigated using 15 commercial HIV seroconversion panels. Panels were selected for short bleeding intervals (generally 7 days). Test results are given in the table in Appendix 6. Results of the bioelisa HIV-1+2 AG/AB test were compared to data from HIV-1 p24 tests and to that of two current HIV-Ag/Ab combination tests: HIV Ag/Ab test-i and HIV Ag/Ab test-ii of different manufacturers. In summary, the bioelisa HIV-1+2 AG/AB test detected 44 samples as reactive (S/CO 1,0), while the CE-marked comparator HIV Ag/Ab combination test I detected only 42 samples as reactive. To enable a comparison for all 15 seroconversion panels, data of the comparator HIV Ag/Ab combination test I were taken from the vendor s data sheet for panel 9030. The bioelisa HIV-1+2 AG/AB test detected HIV infection either in the same bleed as HIV Ag/Ab test-i or one bleed earlier (panels 6240 and 6247). The HIV Ag/Ab test-ii is currently the most sensitive HIV Ag/Ab combination test for which comparative data is available at the PEI Testing Laboratory. The results of eleven panels can be compared to the HIV Ag/Ab test-ii. HIV Ag/Ab test-ii data is not yet available for the panels PRB 967, 969, 971 and 972. In summary, the bioelisa HIV-1+2 AG/AB test detected 34 PEI-IVD Page 8 of 34

bioelisa HIV-1+2 AG/AB samples as reactive while the comparator test detected only 33 samples as reactive. The reactivity of HIV Ag/Ab test-ii on panel member 9034-7 is considered an unspecific reaction. In conclusion, the sensitivity of the Biokit bioelisa HIV-1+2 AG/AB test is comparable to current CE-marked HIV Ag/Ab combination tests with high sensitivity in early infection. Specificity Specificity was assessed using 500 samples from the daily routine of the DRK Frankfurt, Germany. All samples were clearly negative upon the initial test. None of the samples was reactive nor grey zone reactive. Initial and repeat specificity thus were 100%. Mean S/CO value of the 500 samples was 0,23; the median value was 0,22. Minimum S/CO was 0,07 and the maximum S/CO ratio was 0,63 (Fig. 1, Table 6 and Appendix 7). Fig. 1. Distribution Histogram of S/CO Ratios of Blood Donor Samples. Table 6. Specificity Summary S/CO Mean 0,23 Median 0,22 SD 0,04 Mean + 3SD 0,35 Mean - 3SD 0,10 Min 0,07 Max 0,63 SD: Standard deviation PEI-IVD Page 9 of 34

bioelisa HIV-1+2 AG/AB Summary and Conclusions In the study, the clinical sensitivity and specificity of the bioelisa HIV-1+2 AG/AB test was evaluated with 113 anti-hiv-1 positive samples, 4 HIV subtype O and 27 anti-hiv-2 positive samples, 14 HIV-1 p24 antigen subtype samples, 15 commercial seroconversion panels and 500 blood donor samples. Results of the HIV-1 positive samples were compared to those obtained with CE-marked tests used for determining the sample status at the time of sample receipt. For the seroconversion panels two current CE-marked HIV-Ag/Ab combination tests and a sensitive CE-marked Anti-HIV-1/2 test were used for sample characterization. In addition, results from HIV-1 p24 tests and supplemental or confirmatory tests were taken into account for the characterization of the seroconversion panels. Overall sensitivity on the 113 anti-hiv-1 positive samples, on the four HIV-1 subtype O samples and on the 27 anti-hiv-2 positive samples was 100%. S/CO ratios were always high when samples were tested without dilution. There were no results discrepant from the initial sample status. Analytical sensitivity was evaluated using the PEI HIV-1 p24 reference material. The bioelisa HIV-1+2 AG/AB test exhibited a detection limit (9,42 pg/ml) for the PEI HIV-1 p24 reference material which is in a reasonable good range of current assays. Seroconversion panel results were in accordance with the good analytical sensitivity for HIV-1 p24 and demonstrate early detection of HIV infection similar to that of two current CE-marked HIV-Ag/Ab combination tests that exhibit either a good or an excellent sensitivity in early HIV infection. For the investigation of the detection of HIV-1 antigen subtypes 14 samples from the former Nationales Referenzzentrum (NRZ) für Retroviren, Virologisches Institut - Klinische und Molekulare Virologie in Erlangen, Germany were used. These samples constitute supernatants from cell cultures diluted into plasma. Overall, 9 samples were positive, one sample was in the grey zone and two samples showed elevated S/CO ratios (0,83 and 0,88). Sample 14 (subtype O) was negative. Specificity was evaluated on 500 blood donor sera originating from the DRK Hessen-Baden- Württemberg in Frankfurt. All samples tested initially negative and thus initial and repeat specificity was 100 % in this sample collection. Langen, 12/03/2013... (Dr. Sigrid Nick) PEI-IVD Page 10 of 34

bioelisa HIV-1+2 AG/AB Appendix 1 Results of PEI HIV-1 p24 Antigen Dilutions PEI HIV-1 p24 Ag [pg/ml] bioelisa HIV-1+2 Ag/Ab bioelisa HIV-1+2 Ag/Ab bioelisa HIV-1+2 Ag/Ab Lot: J-1912 J-1912 J-1912 B.-No.: 104s/12 104s/12 104s/12 Test date: 24.01.2013 28.01.2013 29.01.2013 S/CO S/CO Mean S/CO S/CO S/CO Mean S/CO S/CO S/CO Mean S/CO Grand Mean [pg/ml] 100,00 6,19 6,59 6,39 7,62 7,16 7,39 6,71 7,09 6,90 50,00 3,23 3,71 3,47 3,97 4,00 3,99 3,74 3,84 3,79 25,00 2,06 2,08 2,07 2,07 2,22 2,14 2,16 2,2 2,18 12,50 1,14 1,15 1,14 1,15 1,22 1,19 1,12 1,19 1,16 6,25 0,70 0,71 0,70 0,73 0,78 0,76 1,09 0,79 0,94 3,13 0,44 0,44 0,44 0,43 0,47 0,45 0,43 0,47 0,45 1,60 0,37 0,39 0,38 0,29 0,41 0,35 0,38 0,36 0,37 Dilution Matrix 0,30 0,40 0,35 0,24 0,35 0,29 0,35 0,38 0,37 Analytical Sensitivity [pg/ml] 10,45 9,81 8,00 9,42 PEI-IVD Page 11 of 34

bioelisa HIV-1+2 AG/AB Appendix 2 Results of Anti-HIV-1 Positive Samples Test: bioelisa HIV-1+2 Ag/Ab Murex HIV Ag/Ab Combination PRISM HIV O PLUS AxSYM HIV Ag/Ab Combo Lots: J-1912 L209510; L223110 69439HN00; 70119HN00 69417LF00 Test dates: 15.01.2013 22.09.2008 27.10.2008 29.09.2008 23.10.2008 13.10.2008 No. W.-No. Date of receipt S/CO S/CO S/CO S/CO 1 113008 16.09.2008 16,85 18,10 n.d. n.d. 2 113013 16.09.2008 16,82 17,49 n.d. n.d. 3 113023 16.09.2008 16,92 17,43 n.d. n.d. 4 113032 16.09.2008 17,45 17,87 n.d. n.d. 5 113064 17.09.2008 18,90 17,41 n.d. n.d. 6 113078 18.09.2008 17,65 16,87 n.d. n.d. 7 113084 18.09.2008 18,97 17,47 n.d. n.d. 8 113093 19.09.2008 17,15 18,63 n.d. n.d. 9 113097 19.09.2008 19,22 18,19 n.d. n.d. 10 113099 19.09.2008 18,84 17,67 n.d. n.d. 11 113100 19.09.2008 19,51 18,98 n.d. n.d. 12 113102 19.09.2008 19,51 > max n.d. n.d. 13 113104 19.09.2008 19,97 > max n.d. n.d. 14 113133 25.09.2008 25,00 n.d. 99,55 22,47 15 113135 25.09.2008 21,25 n.d. 112,64 26,64 16 113136 25.09.2008 17,30 n.d. 95,17 n.d. 17 113137 26.09.2008 18,74 n.d. 104,98 n.d. 18 113138 26.09.2008 21,07 n.d. 107,99 n.d. 19 113139 26.09.2008 20,84 n.d. 90,46 n.d. 20 113140 26.09.2008 23,18 n.d. 111,73 n.d. 21 113141 26.09.2008 19,32 n.d. 120,46 n.d. 22 113142 26.09.2008 21,56 n.d. 113,86 n.d. 23 113143 26.09.2008 24,55 n.d. 123,10 n.d. 24 113144 26.09.2008 17,85 n.d. 103,23 n.d. 25 113145 26.09.2008 19,08 n.d. 109,88 n.d. PEI-IVD Page 12 of 34

bioelisa HIV-1+2 AG/AB Test: bioelisa HIV-1+2 Ag/Ab Murex HIV Ag/Ab Combination PRISM HIV O PLUS AxSYM HIV Ag/Ab Combo 26 113146 26.09.2008 19,81 n.d. 114,31 n.d. 27 113150 26.09.2008 20,57 n.d. 101,45 n.d. 28 113157 30.09.2008 20,50 n.d. n.d. 29,04 29 113167 30.09.2008 19,68 n.d. n.d. 29,30 30 113181 01.10.2008 18,16 n.d. n.d. 10,46 31 113190 02.10.2008 21,94 n.d. n.d. 26,70 32 113192 02.10.2008 17,90 n.d. n.d. 27,42 33 113193 02.10.2008 18,66 n.d. n.d. 28,05 34 113195 02.10.2008 18,73 n.d. n.d. 22,12 35 113199 02.10.2008 19,16 n.d. n.d. 24,96 36 113200 02.10.2008 19,35 n.d. n.d. 20,54 37 113201 02.10.2008 19,73 n.d. n.d. 25,12 38 113202 02.10.2008 19,24 n.d. n.d. 23,59 39 113239 08.10.2008 19,54 n.d. n.d. 57,93 40 113295 10.10.2008 15,99 n.d. n.d. 19,93 41 113297 10.10.2008 15,70 n.d. n.d. 39,35 42 113298 10.10.2008 16,06 n.d. n.d. 51,30 43 113299 10.10.2008 16,58 n.d. n.d. 38,34 44 113300 10.10.2008 18,65 n.d. n.d. 45,49 45 113301 10.10.2008 16,60 n.d. n.d. 59,84 46 113302 10.10.2008 17,20 n.d. n.d. 23,22 47 113303 10.10.2008 15,47 n.d. n.d. 50,42 48 113304 10.10.2008 16,18 n.d. n.d. 24,18 49 113306 10.10.2008 16,39 n.d. n.d. 11,46 50 113307 10.10.2008 18,84 n.d. n.d. 23,39 51 113308 10.10.2008 17,18 n.d. n.d. 21,77 52 113309 10.10.2008 17,56 n.d. n.d. 27,45 53 113335 14.10.2008 17,46 n.d. 118,87 n.d. 54 113338 14.10.2008 18,77 n.d. 120,82 n.d. 55 113479 15.10.2008 15,65 n.d. 133,00 n.d. 56 113494 15.10.2008 17,26 n.d. 129,10 n.d. 57 113508 15.10.2008 16,51 n.d. 153,32 n.d. PEI-IVD Page 13 of 34

bioelisa HIV-1+2 AG/AB Test: bioelisa HIV-1+2 Ag/Ab Murex HIV Ag/Ab Combination PRISM HIV O PLUS AxSYM HIV Ag/Ab Combo 58 113527 17.10.2008 16,96 n.d. 118,96 n.d. 59 113528 17.10.2008 17,12 n.d. 155,63 n.d. 60 113529 17.10.2008 18,24 n.d. 161,20 n.d. 61 113530 17.10.2008 16,89 n.d. 154,65 n.d. 62 113531 17.10.2008 16,96 n.d. 139,94 n.d. 63 113532 17.10.2008 15,48 n.d. 150,22 n.d. 64 113553 17.10.2008 15,83 n.d. 118,51 n.d. 65 113578 22.10.2008 16,38 16,64 n.d. 41,32 66 113579 22.10.2008 16,65 16,27 n.d. 64,81 67 113580 22.10.2008 17,49 16,47 n.d. 65,61 68 113581 22.10.2008 16,82 15,60 n.d. 65,44 69 113583 22.10.2008 16,59 16,35 n.d. 61,73 70 113584 22.10.2008 17,20 16,01 n.d. 56,62 71 113585 22.10.2008 15,38 15,56 n.d. 61,64 72 113587 22.10.2008 20,18 15,78 n.d. 50,45 73 113588 22.10.2008 17,18 17,33 n.d. 66,67 74 113589 22.10.2008 18,22 16,73 n.d. 44,31 75 113590 22.10.2008 17,78 16,55 n.d. 72,00 76 113591 22.10.2008 21,87 17,02 n.d. 53,81 77 113592 22.10.2008 17,70 16,40 n.d. 64,55 78 113593 22.10.2008 24,72 17,79 n.d. 59,48 79 113639 28.10.2008 17,06 n.d. n.d. 16,55 80 113658 29.10.2008 17,59 n.d. n.d. 53,69 81 113671 29.10.2008 17,07 n.d. n.d. 43,53 82 113703 31.10.2008 17,06 n.d. n.d. 69,43 83 113704 31.10.2008 17,13 n.d. n.d. 48,23 84 113709 31.10.2008 20,87 n.d. n.d. 54,38 85 113710 31.10.2008 15,78 n.d. n.d. 52,71 n.d. not determined PEI-IVD Page 14 of 34

bioelisa HIV-1+2 AG/AB Appendix 3 Results of Fresh Anti-HIV-1 Positive Samples bioelisa HIV-1+2 Ag/Ab Murex HIV-1.2.0 bioelisa HIV-1+2 Ag/Ab Architect HIV Ag/Ab Combo Manufacturer: Biokit S.A. DiaSorin Biokit S.A. Abbott Lot: J-1912 D134010 J-1912 15746LI00 Test date: 18.12.2012 18.12.2012 15.01.2013 15.01.2013 No. W.-Nr. Date of Receipt S/CO S/CO S/CO S/CO 1 133339 18.12.2012 16,33 13,87 n.d. n.d. 2 133340 18.12.2012 16,20 13,14 n.d. n.d. 3 133341 18.12.2012 16,64 13,37 n.d. n.d. 4 133342 18.12.2012 16,46 13,63 n.d. n.d. 5 133343 18.12.2012 17,58 12,50 n.d. n.d. 6 133344 18.12.2012 17,13 12,40 n.d. n.d. 7 133345 18.12.2012 19,98 13,02 n.d. n.d. 8 133346 18.12.2012 17,00 12,87 n.d. n.d. 9 133347 18.12.2012 16,76 13,21 n.d. n.d. 10 133348 18.12.2012 18,21 14,39 n.d. n.d. 11 133349 18.12.2012 17,30 14,02 n.d. n.d. 12 133350 18.12.2012 18,44 14,02 n.d. n.d. 13 133351 18.12.2012 19,06 13,75 n.d. n.d. 14 133352 18.12.2012 19,31 13,92 n.d. n.d. 15 133516 15.01.2013 n.d. n.d. 19,77 380,44 16 133517 15.01.2013 n.d. n.d. 18,77 66,51 17 133518 15.01.2013 n.d. n.d. 18,71 429,74 18 133519 15.01.2013 n.d. n.d. 18,98 501,39 19 133520 15.01.2013 n.d. n.d. 21,62 730,09 20 133521 15.01.2013 n.d. n.d. 19,43 416,46 21 133522 15.01.2013 n.d. n.d. 20,85 526,16 22 133523 15.01.2013 n.d. n.d. 18,03 151,26 23 133524 15.01.2013 n.d. n.d. 27,18 288,95 24 133525 15.01.2013 n.d. n.d. 27,19 328,88 25 133526 15.01.2013 n.d. n.d. 21,95 294,71 26 133527 15.01.2013 n.d. n.d. 21,81 638,88 27 133528 15.01.2013 n.d. n.d. 24,43 325,78 28 133529 15.01.2013 n.d. n.d. 20,72 545,44 n.d. not determined PEI-IVD Page 15 of 34

bioelisa HIV-1+2 AG/AB Appendix 4 Results of Anti-HIV-2 Positive Samples Product: Chiron RIBA HIV-1/HIV-2 SIA bioelisa HIV-1+2 Ag/Ab Manufacturer: Novartis Biokit SAU Lot: XA6518 / XA1371 J-1912 Dates of Test 19.12.2007 07.01.2008 04.01.2013 No. Lot No. Supplier ID ADVIA Centaur EHIV Index Genie Multispot HIV 1/2 Sample status as indicated by supplier HIV-1 gp120 HIV-1 gp41 HIV-2 Peptide HIV-1 p31 HIV-1/2 p24 / p26 Sample Status S/CO 1 9212596 10755-16 > 50 HIV-2 HIV-2 positive 1+ 1+ 4+ 4+ 4+ HIV-1/-2 positive 20,45 2 9212611 10770-12 > 50 HIV-2 HIV-2 positive - +/- 4+ 1+ 1+ HIV-2 positive 21,83 3 9212612 10772-13 > 50 HIV-2 HIV-2 positive - 1+ 4+ 4+ 4+ HIV-2 positive 19,06 4 9212614 10774-20 > 50 HIV-2 HIV-2 positive 4+ 4+ 4+ 4+ 4+ HIV-1/-2 positive 20,85 5 9212629 10794-14 > 50 HIV-2 HIV-2 positive - +/- 3+ 1+ 4+ HIV-2 positive 21,35 6 9212631 10798-15 > 50 HIV-2 HIV-2 positive - +/- 3+ 1+ 4+ HIV-2 positive 20,39 7 9212633 10800-17 > 50 HIV-2 HIV-2 positive - 1+ 3+ 4+ 4+ HIV-2 positive 21,34 8 9212635 10804-18 > 50 HIV-2 HIV-2 positive - - 4+ 4+ 4+ HIV-2 positive 18,23 9 9212638 10807-15 > 50 HIV-2 HIV-2 positive - +/- 3+ 4+ 1+ HIV-2 positive 18,92 10 9212639 10808-16 > 50 HIV-2 HIV-2 positive - - 4+ 4+ 3+ HIV-2 positive 20,00 11 9212642 10833-13 > 50 HIV-2 HIV-2 positive +/- +/- 4+ 4+ 4+ HIV-2 positive 19,25 12 9212655 10851-19 > 50 HIV-2 HIV-2 positive - 3+ 4+ 4+ 2+ HIV-2 positive 20,85 13 9212659 10859-10 > 50 HIV-2 HIV-2 positive - 3+ 4+ 4+ 2+ HIV-2 positive 20,12 14 9212665 10869-6 > 50 HIV-2 HIV-2 positive +/- +/- 3+ 3+ 4+ HIV-2 positive 21,13 15 9212666 10870-11 > 50 HIV-2 HIV-2 positive - - 3+ 3+ 3+ HIV-2 positive 20,92 16 9212671 10882-11 > 50 HIV-2 HIV-2 positive - 3+ 4+ 4+ 4+ HIV-2 positive 19,07 17 9212672 10884-13 > 50 HIV-2 HIV-2 positive - - 3+ 3+ 3+ HIV-2 positive 17,48 18 9212674 10887-18 > 50 HIV-2 HIV-2 positive - +/- 4+ 4+ 4+ HIV-2 positive 17,75 19 9212676 10891-16 > 50 HIV-2 HIV-2 positive - - 4+ 4+ +/- HIV-2 positive 17,80 20 9212677 10892-9 > 50 HIV-2 HIV-2 positive 4+ 4+ 3+ 4+ 4+ HIV-1/-2 positive 18,34 21 9212680 10896-14 > 50 HIV-2 HIV-2 positive +/- +/- 4+ 4+ 1+ HIV-2 positive 18,43 PEI-IVD Page 16 of 34

bioelisa HIV-1+2 AG/AB 22 9212682 10902-11 > 50 HIV-2 HIV-2 positive - 2+ 4+ 4+ 1+ HIV-2 positive 18,39 23 9212683 10904-12 > 50 HIV-2 HIV-2 positive - - 3+ 4+ 4+ HIV-2 positive 18,19 24 9212684 10906-23 > 50 HIV-2 HIV-2 positive - - 4+ 4+ 1+ HIV-2 positive 17,54 25 9212687 10909-32 > 50 HIV-2 HIV-2 positive - 1+ 4+ 4+ 4+ HIV-2 positive 18,41 PEI samples PEI HIV-2 reference material 26 (1:1000) HIV-2 positive 5,28/ 5,34/ 4,97 27 W69169 HIV-2 positive 5,42/ 4,16/ 4,38 PEI-IVD Page 17 of 34

bioelisa HIV-1+2 AG/AB Appendix 5. HIV-1 p24 Subtype Panel No. HIV-1 Subtype HIV-1 Subtype (Laboratory ID) Production date (NRZ) bioelisa HIV-1+2 Genetic Systems Test: Ag/Ab Test: HIV-1 Ag HIV Ag/Ab Combo Test date: 11.01.2013 Test date: 17.06.2010 21.06.2010 Viral Load copies/ml s/co Dilution factor (PEI) s/co s/co 1 A A 26.08.2005 126600 0,78 3 1,62 2,39 2 B B 24.04.2008 64100 1,16 1,5 2,09 1,96 3 C C 23.09.2005 72600 1,90 1,5 2,47 3,18 4 D D1 28.10.2005 72000 1,24 1,5 2,46 2,73 5 D D2 28.10.2005 128400 0,91 3 1,82 2,47 6 D D3 11.11.2005 73600 1,29 1,5 2,11 1,78 7 E E1 23.09.2005 190000 0,83 3 2,30 3,72 8 E E2 10.11.2005 100200 1,18 2,5 2,09 3,25 9 F F1 20.01.2006 140400 1,48 3 2,17 1,84 10 F F2 24.07.2009 88800 1,01 2 1,22 1,18 11 G G 24.04.2008 207700 1,42 3 2,11 1,36 12 H H 15.09.2008 124300 0,88 3 2,07 3,32 13 O O1 28.05.2009 624000 1,85 3 8,69 21,12 14 O O2 27.10.2008 135500 0,57 3 0,75 8,21 PEI-IVD Page 18 of 34

bioelisa HIV-1+2 AG/AB Appendix 6. Commercial Anti-HIV-1 Seroconversion Panels No. 1 2 3 Panel Panel member Days since 1st Bleed Manufacturer BIO-RAD Biokit Chiron/ Novartis Test Genetic Systems HIV-1 Ag EIA bioelisa HIV-1+2 Ag/Ab HIV Ag/Ab Combination I HIV Ag/Ab Combination II HIV-1/2 Ab RIBA HIV-1/HIV- 2 SIA HIV RNA copies/ml* s/co s/co s/co s/co s/co HIV1 GP120 PRB 960 1 0 < 50 0,13 0,30 0,25 0,12 0,72 n.a. 2 4 < 50 0,13 0,29 0,24 0,09 0,59 n.a. 3 7 < 50 0,12 0,28 0,25 0,09 0,58 n.a. 4 11 < 50 0,12 0,31 0,29 0,11 0,49 n.a. 5 14 < 50 0,07 0,31 0,26 0,11 0,83 n.a. 6 18 < 50 0,07 0,29 0,29 0,12 0,74 n.a. 7 21 2,9 x 10 2 0,06 0,30 0,25 0,21 0,54 n.a. 8 28 2,8 x 10 7 43,82 15,23 max 461,68 0,28 n.a. 9 30 1,1 x 10 7 46,50 15,05 17,87 556,73 0,54 n.a. PRB 963 1 0 < 50 0,17 0,67 0,26 0,11 0,45 n.d. 2 2 < 50 0,16 0,33 0,26 0,09 0,77 n.d. 3 7 < 50 0,15 0,29 0,26 0,12 0,65 n.d. 4 9 < 50 0,18 0,28 0,27 0,15 0,78 n.d. 5 14 7,8 x 10 3 0,35 0,42 0,37 0,28 0,54 n.d. 6 17 9,6 x 10 4 6,33 5,69 2,90 5,87 0,86 n.d. 7 21 6,2 x 10 5 18,61 13,90 9,33 20,10 3,39 n.d. HIV1 GP41 HIV2 env Peptide HIV1 P31 HIV1/2 P24/26 Results PRB 966 1 0 <50 0,13 0,26 0,29 0,14 0,09 - - - +/- - negative 2 2 <50 0,12 0,25 0,30 0,18 0,15 - - +/- - negative 3 20 <50 0,13 0,21 0,28 0,16 0,18 - - - +/- - negative 4 22 <50 0,17 0,22 0,27 0,13 0,35 - - - +/- - negative 5 30 <50 0,06 0,22 0,27 0,21 0,31 - - - +/- - negative PEI-IVD Page 19 of 34

bioelisa HIV-1+2 AG/AB 4 5 6 7 6 35 3,4x10 2 0,20 0,25 0,27 0,18 0,25 - - - +/- - negative 7 37 1,9x10 3 0,22 0,32 0,30 0,30 0,19 - - - +/- - negative 8 44 2,8x10 5 2,63 4,63 1,90 2,10 0,54 - - - +/- - negative 9 48 4,8x10 4 1,04 5,27 4,02 2,10 2,66 - +/- - +/- - negative 10 51 8,2x10 4 1,39 16,70 15,35 9,62 27,72-3+ - +/- - indeterminate PRB 967 1 0 <75 n.d. 0,25 0,27 n.d. n.d. - - - - - negative 2 3 <75 n.d. 0,32 0,25 n.d. n.d. - - - - - negative 3 7 2171 n.d. 0,53 0,43 n.d. n.d. - - - - - negative 4 17 >500000 n.d. 15,97 >18,8 n.d. n.d. - +/- - - - negative indeterminate 5 19 >500000 n.d. 15,51 max n.d. n.d. - 1+ - - - 6 24 58175 n.d. 16,14 16,45 n.d. n.d. - 3+ - - 2+ positive PRB 969 1 0 <75 n.d. 0,24 0,31 n.d. n.d. - - - - - negative 2 29 <75 n.d. 0,24 0,26 n.d. n.d. - - - - - negative 3 48 <75 n.d. 0,24 0,29 n.d. n.d. - - - - - negative 4 53 <75 n.d. 0,33 0,25 n.d. n.d. - - - - - negative 5 55 103 n.d. 0,37 0,29 n.d. n.d. - - - - - negative 6 61 9797 n.d. 0,44 0,44 n.d. n.d. - - - - - negative 7 63 21979 n.d. 0,57 0,60 n.d. n.d. - - - - - negative 8 70 69598 n.d. 15,94 8,70 n.d. n.d. +/- 2+ - - 1+ positive 9 72 81361 n.d. 16,47 10,26 n.d. n.d. +/- 3+ - - 3+ positive 10 77 11240 n.d. 17,46 7,10 n.d. n.d. 1+ 4+ - - 4+ positive PRB 971 1 0 265 n.d. 0,45 0,29 n.d. n.d. - - - - - negative 2 2 3115 n.d. 0,71 0,40 n.d. n.d. - - - - - negative 3 7 243554 n.d. 27,36 18,50 n.d. n.d. - - - - - negative 4 11 >500000 n.d. 27,37 max n.d. n.d. - - - - - negative PRB 972 1 0 150 n.d. 0,38 0,27 n.d. n.d. - - - - - negative 2 3 160 n.d. 0,33 0,26 n.d. n.d. - - - - - negative 3 11 2000 n.d. 0,31 0,30 n.d. n.d. - - - - - negative 4 14 3500 n.d. 0,46 0,50 n.d. n.d. - - - - - negative PEI-IVD Page 20 of 34

bioelisa HIV-1+2 AG/AB 5 18 22000 n.d. 3,30 1,92 n.d. n.d. - +/- - - - negative 6 21 68000 n.d. 5,18 4,27 n.d. n.d. - +/- - - - negative 8 6240 1 0 0 0,39 VNS 0,23 0,16 0,17 VNS VNS VNS VNS VNS n.d. 2 2 0 0,40 VNS 0,23 0,18 0,17 VNS VNS VNS VNS VNS n.d. 3 7 0 0,37 0,21 0,22 0,06 0,12 - - - - - negative 4 9 0 0,38 0,25 0,26 0,16 0,11 - - - - - negative 5 14 0 0,35 0,25 0,24 0,13 0,14 - - - - - negative 6 16 0 0,35 0,22 0,23 0,11 0,14 - - - - - negative 7 21 36200 1,12 1,83 0,66 0,79 0,18 - - - - - negative 8 23 540000 6,13 12,40 4,26 6,40 0,15 - - - - - negative 9 28 8x10 6 27,72 20,60 18,00 188,95 2,42 - - - - - negative 10 30 10x10 6 28,51 17,69 max 270,68 16,63 - +/- - - - negative indeterminate 11 36 1,6x10 6 5,92 16,73 18,22 47,84 max - 4+ - - +/- 12 45 9x10 4 1,09 17,99 17,56 11,30 max - 4+ - 3+ positive 13 53 n.d. 0,70 19,82 18,26 15,50 max - 4+ - 3+ positive 9 6244 1 0 <400 0,14 0,25 0,47 0,12 0,42 negative 2 5 <400 0,18 0,21 0,25 0,14 0,71 negative 3 7 <400 0,16 0,24 0,26 0,13 0,38 negative 4 12 <400 0,15 0,20 0,31 0,11 0,42 negative 5 13 <400 0,16 0,20 0,24 0,19 0,43 negative 6 14 <400 0,15 0,19 0,27 0,12 0,68 negative 7 16 <400 0,22 0,22 0,22 0,21 0,52 negative 8 19 <400 0,17 0,26 0,24 0,17 0,65 negative 9 21 <400 0,17 0,22 0,23 0,17 0,42 negative 10 22 <400 0,15 0,22 0,37 0,17 0,64 negative 11 25 4550 0,15 0,27 0,30 0,13 0,35 negative 12 26 21000 0,26 0,35 0,32 0,36 0,45 negative 13 28 274000 3,87 2,04 1,38 2,96 0,42 negative 14 33 >750000 39,82 18,43 17,28 68,87 0,90 negative 15 35 >750000 31,93 19,25 17,88 54,90 8,55 negative 10 6247 1 0 <500 0,22 0,28 0,32 0,20 0,07 n.d. n.d. 2 2 <500 0,22 0,26 0,36 0,15 0,08 n.d. n.d. 3 7 <500 0,21 0,30 0,37 0,13 0,07 n.d. n.d. PEI-IVD Page 21 of 34

bioelisa HIV-1+2 AG/AB 4 9 <500 0,22 0,29 0,30 0,17 0,08 n.d. n.d. 5 14 <500 0,20 0,28 0,33 0,15 0,08 n.d. n.d. 6 16 <500 0,18 0,29 0,31 0,18 0,09 n.d. n.d. 7 21 32460 1,31 1,69 0,67 1,40 0,13 n.d. n.d. 8 23 236500 6,49 8,33 3,55 8,14 0,09 n.d. n.d. 9 28 >800000 38,44 16,58 max 166,88 0,08 negative 10 30 >800000 40,29 16,40 max 152,46 0,42 negative 11 6248 1 0 <500 0,24 0,29 0,23 0,07 0,06 negative 2 4 <500 0,22 0,30 0,23 0,11 0,09 negative 3 7 <500 0,20 0,29 0,25 0,12 0,09 negative 4 11 <500 0,20 0,27 0,25 0,09 0,09 negative 5 14 561 0,24 0,35 0,28 0,14 0,12 negative 6 18 74220 3,59 4,55 1,62 3,56 0,09 negative 7 25 >800000 40,66 17,13 18,91 883,52 16,01 negative 12 9010 1 0 <50 0,11 0,28 0,25 0,07 0,12 n.d. 2 2 <50 0,12 0,29 0,26 0,42 0,13 n.d. 3 13 <50 0,18 0,27 0,25 0,08 0,11 n.d. 4 18 <50 0,16 0,24 0,26 0,21 0,12 n.d. 5 20 <50 0,17 0,24 0,25 0,10 0,13 n.d. 6 25 <50 0,18 0,20 0,26 0,09 0,14 n.d. 7 32 <50 0,18 0,21 0,24 0,07 0,14 n.d. 8 34 316 0,20 0,27 0,30 0,08 0,12 n.d. 9 41 146954 5,97 8,16 2,85 5,03 0,12 n.d. 10 45 59647 2,16 13,20 2,27 2,29 1,16 +/- negative indeterminate 11 49 42837 1,88 15,16 4,79 3,14 max 1+ 13 9025 1 0 <50 0,40 0,23 0,21 0,23 0,07 n.d. 2 7 <50 0,38 0,27 0,25 0,35 0,08 n.d. 3 14 <50 0,46 0,29 0,22 0,19 0,07 n.d. 4 23 <50 0,40 0,26 0,22 0,20 0,07 n.d. 5 28 <50 0,36 0,26 0,22 0,22 0,06 n.d. 6 37 <50 0,38 0,27 0,22 0,18 0,10 n.d. 7 58 <50 0,41 0,27 0,22 0,22 0,09 n.d. 8 60 <50 0,37 0,28 0,24 0,22 0,07 n.d. PEI-IVD Page 22 of 34

bioelisa HIV-1+2 AG/AB 9 65 <50 0,36 0,23 0,22 0,20 0,06 n.d. 10 68 <50 0,36 0,26 0,23 0,21 0,07 n.d. 11 85 >500000 11,60 10,24 6,50 10,17 0,08 n.d. 12 91 >500000 30,51 18,58 19,80 140,09 2,17 n.d. 14 9030 1 0 <50 0,17 0,25 0,25* 0,08 0,05 n.d. 2 4 <50 0,17 0,25 0,24 0,10 0,05 n.d. 3 7 <50 0,17 0,28 0,23 0,08 0,05 n.d. 4 11 <50 0,16 0,23 0,24 0,13 0,05 n.d. 5 14 <50 0,18 0,23 0,23 0,12 0,08 n.d. 6 18 <50 0,18 0,27 0,24 0,09 0,14 n.d. 7 21 <50 0,17 0,24 0,24 0,25 0,11 n.d. 8 25 <50 0,16 0,24 0,24 0,08 0,10 n.d. 9 28 <50 0,18 0,25 0,24 0,08 0,11 n.d. 10 33 <50 0,22 0,27 0,23 0,09 0,06 n.d. 11 35 <50 0,22 0,25 0,23 0,08 0,06 n.d. 12 40 85 0,18 0,25 0,23 0,08 0,06 n.d. 13 42 1317 0,21 0,34 0,26 0,05 0,08 n.d. 14 47 238947 8,07 10,09 4,70 8,67 0,09 n.d. 15 49 >500000 18,03 17,62 11,43 22,78 0,56 n.d. 16 54 >500000 26,18 25,20 17,87 60,41 max +/- negative 15 9034 1 0 <50 0,18 0,22 0,34 0,20 0,13 n.d. 2 2 <50 0,19 0,24 0,27 0,38 0,11 n.d. 3 7 <50 0,17 0,23 0,32 0,19 0,10 n.d. 4 9 <50 0,20 0,30 0,22 0,16 0,10 n.d. 5 23 <50 0,23 0,27 0,23 0,12 0,11 n.d. 6 25 <50 0,23 0,26 0,26 0,15 0,12 n.d. 7 32 <50 0,22 0,31 0,27 4,38 0,17 n.d. 8 34 <50 0,20 0,27 0,25 0,13 0,17 n.d. 9 39 <50 0,19 0,28 0,28 0,07 0,14 n.d. 10 41 54 0,17 0,29 0,25 0,06 0,16 n.d. 11 46 123310 2,73 3,72 1,34 3,15 0,10 n.d. 12 50 >500000 27,11 19,66 8,21 46,12 0,24 negative 13 53 >500000 34,20 20,35 17,69 72,89 7,35 negative *Data of HIV Ag/Ab Comnination test I are taken from the vendor s sheet for panel 9030. RNA copies/ml are taken from the panel vendors data sheets; n.d. not determined PEI-IVD Page 23 of 34

bioelisa HIV-1+2 AG/AB Appendix 7: Blood Donor Samples Manufacturer Biokit Test bioelisa HIV-1+2 Ag/Ab Lot no. J-1912 Expiry date 41433 Testing date 23.-25.01.2013 Plate No. W.-No. Sample Barcode No. OD s/co 1 n.a. Blank n.a. 0,025 n.a. n.a. NC n.a. 0,049 n.a. n.a. NC n.a. 0,048 n.a. n.a. NC n.a. 0,052 n.a. n.a. PC1 n.a. 3,077 n.a. n.a. PC2 n.a. 2,040 n.a. n.a. PC Ag n.a. 3,594 n.a. W 133636 Probe 1 70413140703 0,22 W 133637 Probe 2 70413440881 0,21 W 133638 Probe 3 70413440887 0,21 W 133639 Probe 4 70413440875 0,20 W 133640 Probe 5 70413440878 0,21 W 133641 Probe 6 70413140740 0,37 W 133642 Probe 7 70413140721 0,21 W 133643 Probe 8 70413140725 0,21 W 133644 Probe 9 70413140713 0,21 W 133645 Probe 10 70413140708 0,21 W 133646 Probe 11 70413440883 0,21 W 133647 Probe 12 70413440880 0,23 W 133648 Probe 13 70413440868 0,20 W 133649 Probe 14 70413440876 0,21 W 133650 Probe 15 70413440857 0,21 W 133651 Probe 16 70413140739 0,21 W 133652 Probe 17 70413140722 0,22 W 133653 Probe 18 70413140728 0,22 W 133654 Probe 19 70413140711 0,22 W 133655 Probe 20 70413140702 0,23 W 133656 Probe 21 70413440877 0,24 W 133657 Probe 22 70413440884 0,21 W 133658 Probe 23 70413440869 0,22 W 133659 Probe 24 70413440871 0,22 W 133660 Probe 25 70413440856 0,23 W 133661 Probe 26 70413140716 0,21 W 133662 Probe 27 70413140723 0,20 W 133663 Probe 28 70413140715 0,22 W 133664 Probe 29 70413140709 0,21 W 133665 Probe 30 70413140695 0,20 W 133666 Probe 31 70413440879 0,22 W 133667 Probe 32 70413440882 0,21 W 133668 Probe 33 70413440866 0,22 W 133669 Probe 34 70413440874 0,22 W 133670 Probe 35 70413440858 0,22 W 133671 Probe 36 70413140720 0,22 W 133672 Probe 37 70413140724 0,21 W 133673 Probe 38 70413140717 0,19 PEI-IVD Page 24 of 34

bioelisa HIV-1+2 AG/AB W 133674 Probe 39 70413140705 0,21 W 133675 Probe 40 70413140694 0,22 W 133676 Probe 41 70413440885 0,23 W 133677 Probe 42 70413440888 0,25 W 133678 Probe 43 70413440873 0,24 W 133679 Probe 44 70413440872 0,21 W 133680 Probe 45 70413140736 0,23 W 133681 Probe 46 70413140718 0,23 W 133682 Probe 47 70413140727 0,24 W 133683 Probe 48 70413140714 0,24 W 133684 Probe 49 70413140707 0,26 W 133685 Probe 50 70413140696 0,22 W 133686 Probe 51 70413140701 0,21 W 133687 Probe 52 70413440817 0,21 W 133688 Probe 53 70413340151 0,21 W 133689 Probe 54 70413340156 0,21 W 133690 Probe 55 70413340147 0,21 W 133691 Probe 56 70413340143 0,22 W 133692 Probe 57 70413340164 0,24 W 133693 Probe 58 70413340167 0,24 W 133694 Probe 59 70413520378 0,23 W 133695 Probe 60 70413520368 0,22 W 133696 Probe 61 70413140698 0,25 W 133697 Probe 62 70413440822 0,24 W 133698 Probe 63 70413340150 0,24 W 133699 Probe 64 70413340158 0,23 W 133700 Probe 65 70413340152 0,24 W 133701 Probe 66 70413340136 0,26 W 133702 Probe 67 70413340166 0,25 W 133703 Probe 68 70413520400 0,22 W 133704 Probe 69 70413520379 0,21 W 133705 Probe 70 70413520404 0,21 W 133706 Probe 71 70413440778 0,24 W 133707 Probe 72 70413440809 0,22 W 133708 Probe 73 70413340142 0,22 W 133709 Probe 74 70413340163 0,24 W 133710 Probe 75 70413340130 0,29 W 133711 Probe 76 70413340127 0,24 W 133712 Probe 77 70413340178 0,22 W 133713 Probe 78 70413520399 0,28 W 133714 Probe 79 70413520380 0,28 W 133715 Probe 80 70413520393 0,28 W 133716 Probe 81 70413440827 0,24 W 133717 Probe 82 70413440816 0,23 W 133718 Probe 83 70413340145 0,22 W 133719 Probe 84 70413340157 0,20 W 133720 Probe 85 70413340144 0,21 W 133721 Probe 86 70413340138 0,26 W 133722 Probe 87 70413340168 0,21 W 133723 Probe 88 70413520394 0,22 W 133724 Probe 89 70413520356 0,21 2 n.a. Blank n.a. 0,027 n.a. n.a. NC n.a. 0,058 n.a. PEI-IVD Page 25 of 34

bioelisa HIV-1+2 AG/AB n.a. NC n.a. 0,047 n.a. n.a. NC n.a. 0,048 n.a. n.a. PC1 n.a. 3,018 n.a. n.a. PC2 n.a. 2,096 n.a. n.a. PC Ag n.a. 3,611 n.a. W 133725 Probe 90 70413180004 0,23 W 133726 Probe 91 70413440823 0,23 W 133727 Probe 92 70413440811 0,22 W 133728 Probe 93 70413340148 0,22 W 133729 Probe 94 70413340153 0,21 W 133730 Probe 95 70413340128 0,20 W 133731 Probe 96 70413340134 0,21 W 133732 Probe 97 70413520396 0,22 W 133733 Probe 98 70413340176 0,21 W 133734 Probe 99 70413520363 0,22 W 133735 Probe 100 70413180009 0,16 W 133736 Probe 101 70513220140 0,20 W 133737 Probe 102 70513220208 0,21 W 133738 Probe 103 70513220144 0,20 W 133739 Probe 104 70513220206 0,21 W 133740 Probe 105 70513220146 0,07 W 133741 Probe 106 70513220204 0,20 W 133742 Probe 107 70513220193 0,21 W 133743 Probe 108 70513220135 0,23 W 133744 Probe 109 70513220190 0,24 W 133745 Probe 110 70513220134 0,23 W 133746 Probe 111 70513220145 0,22 W 133747 Probe 112 70513220200 0,21 W 133748 Probe 113 70413520397 0,21 W 133749 Probe 114 70513220199 0,23 W 133750 Probe 115 70513220137 0,20 W 133751 Probe 116 70513220196 0,19 W 133752 Probe 117 70513220097 0,20 W 133753 Probe 118 70513220151 0,22 W 133754 Probe 119 70513220092 0,19 W 133755 Probe 120 70513220153 0,24 W 133756 Probe 121 70513220128 0,21 W 133757 Probe 122 70513220185 0,25 W 133758 Probe 123 70513220116 0,21 W 133759 Probe 124 70513220186 0,21 W 133760 Probe 125 70513220113 0,20 W 133761 Probe 126 70513220180 0,36 W 133762 Probe 127 70513220111 0,20 W 133763 Probe 128 70513220162 0,22 W 133764 Probe 129 70513220106 0,20 W 133765 Probe 130 70513220161 0,26 W 133766 Probe 131 70513220118 0,21 W 133767 Probe 132 70513220169 0,25 W 133768 Probe 133 70513220100 0,21 W 133769 Probe 134 70513220166 0,23 W 133770 Probe 135 70513220109 0,24 W 133771 Probe 136 70513220181 0,24 W 133772 Probe 137 70513220115 0,24 PEI-IVD Page 26 of 34

bioelisa HIV-1+2 AG/AB W 133773 Probe 138 70513220171 0,27 W 133774 Probe 139 70513220117 0,23 W 133775 Probe 140 70513220172 0,20 W 133776 Probe 141 70513220093 0,20 W 133777 Probe 142 70513220165 0,20 W 133778 Probe 143 70513220096 0,21 W 133779 Probe 144 70513220163 0,21 W 133780 Probe 145 70513220105 0,22 W 133781 Probe 146 70513220183 0,23 W 133782 Probe 147 70513220132 0,24 W 133783 Probe 148 70513220187 0,24 W 133784 Probe 149 70513220126 0,21 W 133785 Probe 150 70513220173 0,22 W 133786 Probe 151 70513220188 0,47 W 133787 Probe 152 70513220167 0,24 W 133788 Probe 153 70513220138 0,24 W 133789 Probe 154 70513220197 0,24 W 133790 Probe 155 70513220142 0,26 W 133791 Probe 156 70513220194 0,22 W 133792 Probe 157 70513220141 0,21 W 133793 Probe 158 70513220237 0,23 W 133794 Probe 159 70513220207 0,23 W 133795 Probe 160 70413140742 0,22 W 133796 Probe 161 70513220203 0,22 W 133797 Probe 162 70513220121 0,26 W 133798 Probe 163 70513220148 0,21 W 133799 Probe 164 70513220099 0,22 W 133800 Probe 165 70513220155 0,23 W 133801 Probe 166 70513220107 0,23 W 133802 Probe 167 70413440836 0,24 W 133803 Probe 168 70513220249 0,23 W 133804 Probe 169 70513220254 0,23 W 133805 Probe 170 70413140741 0,26 W 133806 Probe 171 70513220198 0,21 W 133807 Probe 172 70513220108 0,21 W 133808 Probe 173 70513220160 0,21 W 133809 Probe 174 70513220110 0,24 W 133810 Probe 175 70513220154 0,21 W 133811 Probe 176 70513220112 0,22 W 133812 Probe 177 70513220156 0,23 W 133813 Probe 178 70513220217 0,23 3 n.a. Blank n.a. 0,020 n.a. n.a. NC n.a. 0,050 n.a. n.a. NC n.a. 0,046 n.a. n.a. NC n.a. 0,050 n.a. n.a. PC1 n.a. 3,247 n.a. n.a. PC2 n.a. 2,205 n.a. n.a. PC Ag n.a. 3,769 n.a. W 133814 Probe 179 70513220238 0,22 W 133815 Probe 180 70413140744 0,21 W 133816 Probe 181 70513220189 0,20 W 133817 Probe 182 70513220114 0,22 W 133818 Probe 183 70513220182 0,20 PEI-IVD Page 27 of 34

bioelisa HIV-1+2 AG/AB W 133819 Probe 184 70513220102 0,19 W 133820 Probe 185 70513220174 0,20 W 133821 Probe 186 70513220122 0,19 W 133822 Probe 187 70513220147 0,22 W 133823 Probe 188 70513220250 0,19 W 133824 Probe 189 70513220232 0,20 W 133825 Probe 190 70413140745 0,20 W 133826 Probe 191 70513220176 0,23 W 133827 Probe 192 70513220133 0,19 W 133828 Probe 193 70513220184 0,18 W 133829 Probe 194 70513220125 0,19 W 133830 Probe 195 70513220195 0,31 W 133831 Probe 196 70513220127 0,22 W 133832 Probe 197 70513220216 0,25 W 133833 Probe 198 70513220201 0,22 W 133834 Probe 199 70513220220 0,24 W 133835 Probe 200 70413140746 0,22 W 133836 Probe 201 70413140704 0,21 W 133837 Probe 202 70413440849 0,22 W 133838 Probe 203 70413440839 0,23 W 133839 Probe 204 70413440855 0,21 W 133840 Probe 205 70413440850 0,18 W 133841 Probe 206 70413440870 0,19 W 133842 Probe 207 70413440859 0,19 W 133843 Probe 208 70513220230 0,18 W 133844 Probe 209 70513220234 0,19 W 133845 Probe 210 70513220104 0,22 W 133846 Probe 211 70413140697 0,24 W 133847 Probe 212 70413440843 0,63 W 133848 Probe 213 70413440837 0,23 W 133849 Probe 214 70413440861 0,24 W 133850 Probe 215 70413440853 0,23 W 133851 Probe 216 70413440864 0,21 W 133852 Probe 217 70413440862 0,22 W 133853 Probe 218 70513220239 0,22 W 133854 Probe 219 70513220227 0,24 W 133855 Probe 220 70513220129 0,23 W 133856 Probe 221 70413140700 0,25 W 133857 Probe 222 70413440840 0,24 W 133858 Probe 223 70413440841 0,29 W 133859 Probe 224 70413440846 0,24 W 133860 Probe 225 70413440854 0,26 W 133861 Probe 226 70413440865 0,25 W 133862 Probe 227 70513220260 0,25 W 133863 Probe 228 70513220246 0,24 W 133864 Probe 229 70513220258 0,23 W 133865 Probe 230 70513220152 0,25 W 133866 Probe 231 70413140699 0,25 W 133867 Probe 232 70413440844 0,22 W 133868 Probe 233 70413440838 0,23 W 133869 Probe 234 70413440860 0,22 W 133870 Probe 235 70413440848 0,26 W 133871 Probe 236 70413440863 0,27 PEI-IVD Page 28 of 34

bioelisa HIV-1+2 AG/AB W 133872 Probe 237 70513220214 0,58 W 133873 Probe 238 70513220225 0,27 W 133874 Probe 239 70513220101 0,24 W 133875 Probe 240 70513220150 0,25 W 133876 Probe 241 70413440779 0,24 W 133877 Probe 242 70413440842 0,22 W 133878 Probe 243 70413440829 0,24 W 133879 Probe 244 70413440851 0,23 W 133880 Probe 245 70413440847 0,24 W 133881 Probe 246 70413440867 0,21 W 133882 Probe 247 70513220221 0,25 W 133883 Probe 248 70513220212 0,24 W 133884 Probe 249 70513220103 0,22 W 133885 Probe 250 70513220170 0,24 W 133886 Probe 251 70513220175 0,23 W 133887 Probe 252 70513220120 0,22 W 133888 Probe 253 70513220228 0,24 W 133889 Probe 254 70513220218 0,23 W 133890 Probe 255 70413520390 0,24 W 133891 Probe 256 70513220219 0,23 W 133892 Probe 257 70513220224 0,28 W 133893 Probe 258 70513220211 0,22 W 133894 Probe 259 70413140738 0,25 W 133895 Probe 260 70413140752 0,25 W 133896 Probe 261 70513220191 0,27 W 133897 Probe 262 70513220131 0,22 W 133898 Probe 263 70513220236 0,23 W 133899 Probe 264 70513220235 0,21 W 133900 Probe 265 70513220233 0,22 W 133901 Probe 266 70513220231 0,23 W 133902 Probe 267 70513220229 0,21 4 n.a. Blank n.a. 0,028 n.a. n.a. NC n.a. 0,056 n.a. n.a. NC n.a. 0,045 n.a. n.a. NC n.a. 0,046 n.a. n.a. PC1 n.a. 3,381 n.a. n.a. PC2 n.a. 2,537 n.a. n.a. PC Ag n.a. 4,923 n.a. W 133903 Probe 268 70513220240 0,38 W 133904 Probe 269 70413140735 0,22 W 133905 Probe 270 70413140751 0,22 W 133906 Probe 271 70513220205 0,21 W 133907 Probe 272 70513220149 0,23 W 133908 Probe 273 70513220241 0,22 W 133909 Probe 274 70513220252 0,23 W 133910 Probe 275 70513220244 0,22 W 133911 Probe 276 70513220251 0,27 W 133912 Probe 277 70513220243 0,27 W 133913 Probe 278 70513220226 0,24 W 133914 Probe 279 70413140732 0,25 W 133915 Probe 280 70413140748 0,24 W 133916 Probe 281 70513220095 0,23 W 133917 Probe 282 70513220164 0,22 PEI-IVD Page 29 of 34

bioelisa HIV-1+2 AG/AB W 133918 Probe 283 70513220256 0,22 W 133919 Probe 284 70513220259 0,27 W 133920 Probe 285 70513220253 0,23 W 133921 Probe 286 70513220257 0,22 W 133922 Probe 287 70513220255 0,25 W 133923 Probe 288 70413140737 0,23 W 133924 Probe 289 70413140734 0,22 W 133925 Probe 290 70413140747 0,26 W 133926 Probe 291 70513220094 0,23 W 133927 Probe 292 70513220168 0,29 W 133928 Probe 293 70513220223 0,23 W 133929 Probe 294 70513220215 0,23 W 133930 Probe 295 70513220210 0,24 W 133931 Probe 296 70513220192 0,22 W 133932 Probe 297 70513220209 0,24 W 133933 Probe 298 70413140733 0,24 W 133934 Probe 299 70413140730 0,24 W 133935 Probe 300 70413140743 0,27 W 133936 Probe 301 70413440781 0,25 W 133937 Probe 302 70413180003 0,26 W 133938 Probe 303 70413440774 0,24 W 133939 Probe 304 70413440789 0,23 W 133940 Probe 305 70413440783 0,25 W 133941 Probe 306 70413440794 0,22 W 133942 Probe 307 70413440799 0,24 W 133943 Probe 308 70413520415 0,25 W 133944 Probe 309 70413520459 0,27 W 133945 Probe 310 70413100625 0,26 W 133946 Probe 311 70413440785 0,27 W 133947 Probe 312 70413180011 0,24 W 133948 Probe 313 70413440771 0,27 W 133949 Probe 314 70413440788 0,27 W 133950 Probe 315 70413440784 0,23 W 133951 Probe 316 70413440812 0,27 W 133952 Probe 317 70413440798 0,25 W 133953 Probe 318 70413520407 0,22 W 133954 Probe 319 70413520464 0,23 W 133955 Probe 320 70413100623 0,23 W 133956 Probe 321 70413440776 0,22 W 133957 Probe 322 70413180016 0,24 W 133958 Probe 323 70413440772 0,26 W 133959 Probe 324 70413440782 0,25 W 133960 Probe 325 70413440780 0,27 W 133961 Probe 326 70413440805 0,25 W 133962 Probe 327 70413520435 0,23 W 133963 Probe 328 70413520411 0,56 W 133964 Probe 329 70413520463 0,25 W 133965 Probe 330 70413100617 0,24 W 133966 Probe 331 70413440777 0,23 W 133967 Probe 332 70413180015 0,23 W 133968 Probe 333 70413440801 0,24 W 133969 Probe 334 70413440786 0,29 W 133970 Probe 335 70413440790 0,24 PEI-IVD Page 30 of 34

bioelisa HIV-1+2 AG/AB W 133971 Probe 336 70413440800 0,24 W 133972 Probe 337 70413520427 0,24 W 133973 Probe 338 70413520465 0,22 W 133974 Probe 339 70413520460 0,22 W 133975 Probe 340 70413100607 0,24 W 133976 Probe 341 70413440775 0,24 W 133977 Probe 342 70413440773 0,27 W 133978 Probe 343 70413440793 0,24 W 133979 Probe 344 70413440797 0,27 W 133980 Probe 345 70413440814 0,27 W 133981 Probe 346 70413440806 0,27 W 133982 Probe 347 70413520424 0,27 W 133983 Probe 348 70413520458 0,26 W 133984 Probe 349 70413100626 0,24 W 133985 Probe 350 70413100619 0,22 W 133986 Probe 351 70413100612 0,24 W 133987 Probe 352 70413100620 0,24 W 133988 Probe 353 70413520429 0,24 W 133989 Probe 354 70413520446 0,23 W 133990 Probe 355 70413520421 0,22 W 133991 Probe 356 70413520456 0,22 5 n.a. Blank n.a. 0,022 n.a. n.a. NC n.a. 0,047 n.a. n.a. NC n.a. 0,047 n.a. n.a. NC n.a. 0,043 n.a. n.a. PC1 n.a. 2,995 n.a. n.a. PC2 n.a. 2,228 n.a. n.a. PC Ag n.a. 3,488 n.a. W 133992 Probe 357 70413520423 0,20 W 133993 Probe 358 70413520452 0,22 W 133994 Probe 359 70413520405 0,18 W 133995 Probe 360 70413520387 0,22 W 133996 Probe 361 70413100616 0,20 W 133997 Probe 362 70413100621 0,17 W 133998 Probe 363 70413520416 0,20 W 133999 Probe 364 70413520451 0,23 W 134000 Probe 365 70413520422 0,20 W 134001 Probe 366 70413520439 0,19 W 134002 Probe 367 70413520417 0,19 W 134003 Probe 368 70413520447 0,19 W 134004 Probe 369 70413520401 0,18 W 134005 Probe 370 70413520369 0,17 W 134006 Probe 371 70413100614 0,18 W 134007 Probe 372 70413100593 0,23 W 134008 Probe 373 70413520413 0,20 W 134009 Probe 374 70413520444 0,19 W 134010 Probe 375 70413520414 0,18 W 134011 Probe 376 70413520436 0,21 W 134012 Probe 377 70413520419 0,18 W 134013 Probe 378 70413520430 0,22 W 134014 Probe 379 70413520383 0,20 W 134015 Probe 380 70413520319 0,19 W 134016 Probe 381 70413100618 0,20 PEI-IVD Page 31 of 34

bioelisa HIV-1+2 AG/AB W 134017 Probe 382 70413100591 0,19 W 134018 Probe 383 70413520406 0,18 W 134019 Probe 384 70413520437 0,20 W 134020 Probe 385 70413520412 0,19 W 134021 Probe 386 70413520434 0,19 W 134022 Probe 387 70413520410 0,20 W 134023 Probe 388 70413520372 0,23 W 134024 Probe 389 70413520377 0,23 W 134025 Probe 390 70413520360 0,20 W 134026 Probe 391 70413100615 0,20 W 134027 Probe 392 70413100596 0,20 W 134028 Probe 393 70413520454 0,18 W 134029 Probe 394 70413520426 0,20 W 134030 Probe 395 70413520432 0,20 W 134031 Probe 396 70413520425 0,23 W 134032 Probe 397 70413520457 0,20 W 134033 Probe 398 70413520364 0,20 W 134034 Probe 399 70413520386 0,22 W 134035 Probe 400 70413520351 0,21 W 134036 Probe 401 70413520304 0,20 W 134037 Probe 402 70413520334 0,21 W 134038 Probe 403 70413520300 0,22 W 134039 Probe 404 70413520338 0,25 W 134040 Probe 405 70413520309 0,22 W 134041 Probe 406 70413180008 0,19 W 134042 Probe 407 70413520336 0,22 W 134043 Probe 408 70413520307 0,19 W 134044 Probe 409 70413520332 0,18 W 134045 Probe 410 70413520299 0,27 W 134046 Probe 411 70413520333 0,19 W 134047 Probe 412 70413520326 0,19 W 134048 Probe 413 70413520375 0,19 W 134049 Probe 414 70413520329 0,22 W 134050 Probe 415 70413180007 0,23 W 134051 Probe 416 70413180005 0,32 W 134052 Probe 417 70413520316 0,21 W 134053 Probe 418 70413520301 0,21 W 134054 Probe 419 70413520327 0,20 W 134055 Probe 420 70413520310 0,20 W 134056 Probe 421 70413520344 0,20 W 134057 Probe 422 70413520303 0,21 W 134058 Probe 423 70413520346 0,21 W 134059 Probe 424 70413520317 0,20 W 134060 Probe 425 70413180012 0,20 W 134061 Probe 426 70413180002 0,20 W 134062 Probe 427 70413520330 0,19 W 134063 Probe 428 70413520349 0,19 W 134064 Probe 429 70413520324 0,20 W 134065 Probe 430 70413520339 0,20 W 134066 Probe 431 70413520337 0,27 W 134067 Probe 432 70413520305 0,24 W 134068 Probe 433 70413520343 0,20 W 134069 Probe 434 70413520302 0,24 PEI-IVD Page 32 of 34

bioelisa HIV-1+2 AG/AB W 134070 Probe 435 70413180010 0,21 W 134071 Probe 436 70413520347 0,20 W 134072 Probe 437 70413520320 0,20 W 134073 Probe 438 70413520352 0,25 W 134074 Probe 439 70413520311 0,27 W 134075 Probe 440 70413520342 0,22 W 134076 Probe 441 70413520341 0,22 W 134077 Probe 442 70413520306 0,21 W 134078 Probe 443 70413520348 0,20 W 134079 Probe 444 70413520313 0,23 W 134080 Probe 445 70413520373 0,21 5 n.a. Blank n.a. 0,019 n.a. n.a. NC n.a. 0,047 n.a. n.a. NC n.a. 0,041 n.a. n.a. NC n.a. 0,042 n.a. n.a. PC1 n.a. 3,213 n.a. n.a. PC2 n.a. 2,677 n.a. n.a. PC Ag n.a. 3,602 n.a. W 134081 Probe 446 70413520350 0,27 W 134082 Probe 447 70413520314 0,22 W 134083 Probe 448 70413520325 0,20 W 134084 Probe 449 70413520323 0,23 W 134085 Probe 450 70413520345 0,22 W 134086 Probe 451 70413520322 0,22 W 134087 Probe 452 70413520308 0,23 W 134088 Probe 453 70413180077 0,22 W 134089 Probe 454 70413180090 0,30 W 134090 Probe 455 70413180085 0,18 W 134091 Probe 456 70413460931 0,19 W 134092 Probe 457 70413460928 0,18 W 134093 Probe 458 70413520441 0,19 W 134094 Probe 459 70413520409 0,19 W 134095 Probe 460 70413520440 0,19 W 134096 Probe 461 70413520335 0,20 W 134097 Probe 462 70413520370 0,21 W 134098 Probe 463 70413180073 0,22 W 134099 Probe 464 70413180089 0,20 W 134100 Probe 465 70413180084 0,23 W 134101 Probe 466 70413460918 0,24 W 134102 Probe 467 70413460914 0,20 W 134103 Probe 468 70413520445 0,21 W 134104 Probe 469 70413520408 0,19 W 134105 Probe 470 70413520438 0,23 W 134106 Probe 471 70413520328 0,19 W 134107 Probe 472 70413180080 0,20 W 134108 Probe 473 70413180069 0,21 W 134109 Probe 474 70413180087 0,24 W 134110 Probe 475 70413180079 0,19 W 134111 Probe 476 70413460926 0,23 W 134112 Probe 477 70413460906 0,22 W 134113 Probe 478 70413520448 0,29 W 134114 Probe 479 70413520362 0,19 W 134115 Probe 480 70413520442 0,20 PEI-IVD Page 33 of 34

bioelisa HIV-1+2 AG/AB n.a. not applicable W 134116 Probe 481 70413520315 0,21 W 134117 Probe 482 70413180075 0,23 W 134118 Probe 483 70413460934 0,19 W 134119 Probe 484 70413180088 0,21 W 134120 Probe 485 70413460932 0,20 W 134121 Probe 486 70413460925 0,27 W 134122 Probe 487 70413520359 0,23 W 134123 Probe 488 70413520450 0,26 W 134124 Probe 489 70413520420 0,23 W 134125 Probe 490 70413520449 0,23 W 134126 Probe 491 70413520312 0,31 W 134127 Probe 492 70413180078 0,24 W 134128 Probe 493 70413460936 0,23 W 134129 Probe 494 70413180086 0,22 W 134130 Probe 495 70413460929 0,22 W 134131 Probe 496 70413460930 0,22 W 134132 Probe 497 70413520431 0,23 W 134133 Probe 498 70413520355 0,23 W 134134 Probe 499 70413520428 0,23 W 134135 Probe 500 70413520455 0,24 PEI-IVD Page 34 of 34