Global Rapid Diagnostic Leader SD

1. Global Rapid Diagnostic Leader SD

2. RAT (Rapid Antigen Test) SD BIOLINE Influenza Antigen (type A & B)SD BIOLINE Influenza H5 (A/H5N1)

3. SD BIOLINE Influenza Antigen Kit SD BIOLINE Influenza Antigen rapid test is a chromatographic immunoassay for the differential and qualitative detection of influenza virus type A and type B antigens directly from nasal swab specimens, throat swab specimens, nasal/nasopharyngeal aspiratespecimens.

4. Test Principle & Major active ingredient - Detection by Direct Sandwich System mouse monoclonal anti-influenza A/B-gold conjugate Gold Gold Gold Flu A Ag Flu B Ag in specimen in specimen mouse monoclonal anti-influenza B mouse monoclonal anti-influenza A C line B line(Influenza B type) A line(Influenza A type) • Target materials : Influenza antigen (type Aband B) • in respiratory specimen

5. Test Results Appearance Negative Influenza type A Influenza type B positive positive

6. Test procedure & Interpretation • Transfer assay diluent into test tube (approximately 300uL). • Insert the patient swab sample into the test tube and mix well. • Place the Test Strip into the test tube. • Read result at 10 ~ 15 minutes.

7. Sample collection & Test procedure 2. Diluent 300 uL 1. Nasal swab

8. SD BIOLINE Influenza Antigen Kit Sample collection & Test procedure 4. Discard the swab. 3. Extraction

9. SD BIOLINE Influenza Antigen Kit Sample collection & Test procedure 6. Migration 5. Insert test strip into tube.

10. Interpretation of Test Results A/H1N1 A/H3N2 B 3+ 2+ 1+ 3+ 2+ 1+ 3+ 2+ 1+ Neg.

11. Comparison Table (Influenza RAT) Q SD BD

12. Sensitivity per the virus type of strains

13. Sensitivity per the virus type of strains

14. Comparison data vs. RAT competitor Detection Limit of each virus type

15. Clinical Evaluation Studies The performance of the SD BIOLINE Influenza antigen test was compared to cell culture or RT-PCR methods in a multi-center field clinical study. In this multi-center, a combination of nasal swabs and throat swabs specimens were collected from total 341 patients. *Test Period ; Oct.2003 ~ May 2005 *Test Region ; Korea & Overseas Vietnam (H5) HongKong Thailand, etc

16. Clinical Evaluation Studies Summary of Clinical Evaluation Data

17. Clinical evaluation studies results performed at National Institute of Hygiene and Epidemiology, Vietnam (including H5 Human cases) *Test period : Dec.2004 ~ Dec.2005

18. Competitiveness of SD BIOLINE Influenza Antigen test Why should you use “SD BIOLINE Influenza Antigen Rapid Test”? SD kit can cover all influenza virus type. SD kit can detect AIV infection in Human (A/H5N1). SD kit can differentiate influenza virus type A and B SD kit is very useful as POCT (point of care testing), on-site test.

19. SD BIOLINE Influenza H5 kit SD BIOLINE Influenza H5 rapid test is a chromatographic immunoassay for the qualitative detection of hemagglutinin H5 antigen of avian influenza (AI)virus directly from nasal swab/aspirate specimens, throat swab specimens, tracheal aspirate specimens on AI virus human infection cases.

20. Test Principle & Major active ingredient - Detection by Direct Sandwich System mouse monoclonal anti-influenza H5-gold conjugate Gold Gold Flu H5 Ag in specimen mouse monoclonal anti-influenza H5 C line T line (Influenza A/H5) • Target materials : Hemagglutinin H5 antigen • of avian influenza (AI) virus in respiratory specimen

21. specimen collection & preparation • Nasal swab • Nasal aspirate • Throat swab • Tracheal aspirate (Especially, test results with the tracheal aspirate specimens would be recommendable, because the majority of AIV may exist at the tracheal position.)

22. specimen collection & preparation • If the specimen should be tested later, swab specimens should be placed into 1 ~ 2 ml of transport media or saline by direct extraction. • Transport media : Use of the following transport media has been tested and found to be compatible with SD BIOLINE Influenza Antigen test. • Saline EMEM+1%BSA • PBS EMEM+0.5%BSA • PBS+0.5%BSA Trypticase soy Broth+0.5%BSA • PBS+0.5%Gelatin Trypticase soy Broth+0.5% gelatin

23. Special Precautions • on Specimen collection & preparation • Use of fresh sample is very important for getting accurate results. • The specimen which have ever been thawed after freezing may results in decreased test sensitivity from the deterioration or modification of H5 antigen.So,repeated freezing and thawing of specimen must not be used for this test. • Finally, to get accurate results, specimen should be tested as soon as possible after collection.

24. Test procedure & Interpretation • ; nasal swab, throat swab • Transfer assay diluent into test tube (approximately 300uL). • Insert the patient swab sample into the test tube and mix well. • Place the Test Strip into the test tube. • Read result at 10 ~ 15 minutes.

25. Test procedure & Interpretation ; nasal aspirate, tracheal aspirate • Pipet 100uL of assay diluent and 100uL of aspirate specimen into test tube. And mix well. • Place the Test Strip into the test tube. • Read result at 10 ~ 15 minutes.

26. Test Results Appearance ; Influenza A/H5 positive

27. SD BIOLINE Influenza H5 Limit of Detection : H5N1 (*)minimum detectable level With H5N1 (ES/03) : virus titer 27 HA unit MDL(*) 32HAU 16HAU 8HAU 4HAU 2HAU 1HAU +++ +++ ++ ++ + +

28. SD BIOLINE Influenza H5 Evaluation data on AIV H5 antigen

29. SD BIOLINE Influenza H5 Evaluation data on AIV H5 antigen

30. SD BIOLINE Influenza H5 Evaluation data on AIV H5 antigen

31. SD BIOLINE Influenza H5 Evaluation data on AIV H5 antigen • Case 1 : • At the WHO Collaborating Centre for Reference and Research on Influenza • Dr. Aeron Hurt • Address : 45 Poplar Rd,Parkville, Victoria 3052, Australia • SD H5 kit could detect as below; • A/Chicken/Vietnam #8 (-irradiated) – A(H5N1) – Egg grown • A/Duck/Vietnam #7A (-irradiated) – A(H5N1) – Egg grown • A/?/Vietnam #Pooled (-irradiated) – A(H5N1) – Egg grown • Case 2 : • At the Avian Section, Diagnostic Virology Laboratory National Veterinary Services Laboratories, APHIS, USDA • Dr. Janice C. Pedersen, MicrobiologistSD H5 kit could detect as below; • TY/MN H5N2 10-1

32. SD BIOLINE Influenza H5 Evaluation data on AIV H5 antigen • Case 3 : • At the ISTITUTO ZOOPROFILATTICO SPERIMENTALE DELLA • LOMBARDIA E DELL’EMILIA ROMAGNA “BRUNO UBERTINI” • (ENTE SANITARIO DI DIRITTO PUBBLICO) • Address : Via Bianchi, 9, 25124 BRESCIA, ITALY • SD H5 kit could detect as below; • A/ty/It/90302/05/H5N2 (LPAI) - 104,5EID50/ml as detection limit. • Case 4 : At the OIE AIV Reference Laboratory Clinic for bird, reptile, amphibia and fishes Universita"t Giessen, Germany • Justus-Liebig-University Giessen Frankfurter, Germany • Performed by Prof. Dr. E. F. Kaleta, • SD H5 kit could detect as below; • H5Nx: Cell culture supernatant fluids containing H5 virus (H5N2, H5N9, H5N1) • were cpe pos. in chick embryo cell cultures up to 10 to minus three.

33. <Step 1> Surveillance on Central Hospital around region of outbreak of AIV Review & Collection of hospitalized records Collection of blood or respiratory specimen from possible case or not-caused ARDS Laboratory diagnosis at CDC, Dept. Of infectious disease control Poultry AIV – human infection Surveillance of Epidemiology case study by Korea CDC

34. <Step 2> If, detection of AIV_human infection possible case, you should work on enlarged surveillance with all general hospital adjacent to the region of outbreak of AIV. Poultry AIV – human infection Surveillance of Epidemiology case study by Korea CDC

35. Virus isolation in tissue culture cells, or virus inoculation into egg Viral nucleic acid : RT-PCR Hemagglutination Inhibition test (HI) Neutralization (NT) Infectious virus ELISA: recombinant antigen Western blot recombinant antigen Diagnosis of AIV infection to Human

36. Diagnosis of AIV infection to Human Sample collection

37. Diagnosis Flow chart Specimen collection Respiratory specimen Serum First, by Rapid Antigen Testing HI Participates in WHO/CDC programs for flu surveillance RT-PCR / Virus culture NT

38. Respiratory specimen In case of emergency that happen to outbreak of AI in the poultry, SD BIOLINE Influenza H5 test should be used directly. RAT by SD BIOLINE Influenza Ag test Type A positive Type B positive RAT by SD BIOLINE Influenza H5 test Treatment or Confirm test by RT-PCR/viral culture H5 positive negative Treatment or Confirm test by RT-PCR/viral culture Send Samples to Reference Laboratory that participates in WHO/CDC programs for flu/H5 surveillance, immediately Applications of Influenza Rapid kits on Avian Influenza diagnosis programs Negative • **WHO recommendations on the use of rapid testing for influenza diagnosis ; • Influenza surveillance should be used to guide the optimal use of RAT (rapid antigen test). • At the beginning of the influenza season or an influenza outbreak, RAT may influence on clinical decisions and contribute to clinical awareness.

39. SD BIOLINE Influenza H5 Advantages as an aid of H5 diagnosis • For the screening test, the Sole H5 Avian Influenza Antigen Rapid Kit in the world • Accurate detection on H5N1 samples isolated from Human infection cases, with High Sensitivity and Specificity • With same kits, we’ve got lots of AI positive cases through HPAI (H5N1~H5N9) detection in the farm, market, or laboratory by use of feces or cloacal swab as veterinary use. • Easy assay procedure: No technical expert and No equipment is required • Labor & Time saving because of fast results

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