1. Avian Influenza ELISA Test Comparison

2. COMPARATIVE TEST OF 152 EXPERIMENTAL AND NATURAL INFECTED CHICKEN AND TURKEY INFLUENZA “A”- POSITIVE SERA BY USING THREE DIFFERENT ELISAs AND INHIBITION OF HAEMAGGLUTINATION TEST

3. Comparison Testing: Affinitech’s Avian Influenza Virus Antibody Test Kit (Indirect method) Another commercially available Avian Influenza Virus Antibody Test Kit (Indirect method) ELISA HB65 method (Boer et al.,1990, Arch Virol, 115: 47-61) modified by IZT (ISTITUTO ZOOPROFILATTICO SPERIMENTALE) (Competitive ELISA using Mab65 on the plate) and the OIE and CE reference test i.e. inhibition of haemagglutination (HI).

4. The tested sera included: 35 samples of natural infected H7N1 turkey sera from two intensive farms 25 samples of natural infected H7N3 turkey sera from three different farms 40 samples of natural infected H7N3 chicken sera from three farms identified - 12 samples of H7N1 chickens sera from vaccinated birds Internal reference sera (either experimental and field sera with known titre determined after repeated tests) Experimentallly infected SPF chicken with H7N3, H3N2, H10N7, H6N2, H5N2, H1N1 and H9N2 Experimental infected SPF chicken with Newcastle disease virus (NDV) Negative SPF chicken Natural infected H7N3 turkey 20 samples of field negative chicken sera from two farms 10 samples of field negative turkey sera from one farm

5. COMPARATIVE RESULTS The following tables reports the results of tested sera using the three different ELISAs and the HI test. The interpretation values are the following:

6. Natural Infected Field Sera

7. Natural Infected Field Sera (continued)

12. Vaccinated Chickens’ Sera

13. Internal Reference Sera

14. Internal Reference Sera (continued)

15. Negative Field Sera

16. Negative Field Sera (continued)

17. Natural Infected Sera

18. Vaccination Produced Sera

19. Negative Field Sera

20. Negative Field Sera (continued)

22. Statistical Data

23. Experimentally Infected USDA Reference Panel for Avian Influenza

24. S/P Ratio V. Agar Gel Precipitation

25. References An Enzyme-Linked Immunosorbent Assay for the Detection of Antibody Against Avian Influenza Virus by D.B. Snyder, W.W. Marquardt, F.S. Yancy and P.K. Savage, Avian Diseases 29: 136-144 Development of Enzyme-Linked Immunosorbent Assay with Nucleoprotein as Antigen for Detectin of Antibodies to Avian Influenza Virus by Meilin Jin, Guihua Wang, Ruihua Zhang, Siting Zhao, Hangchao Li, Yadi Tan and Huanchun Chen, Avian Diseases 48:870-878, 2004 An ELISA for detection of antibodies against influenza A nucleoprotein in humans and various animal species by G.F. Boer, W. Back and A.D.M.E. Osterhaus, Archives of Virology (1990) 115: 47-61

26. Special Thanks to Dr. P. Cordioli with: ISTITUTO ZOOPROFILATTICO SPERIMENTALE DELLA LOMBARDIA E DELL'EMILIA ROMAGNA “B. UBERTINI” (ENTE SANITARIO DI DIRITTO PUBBLICO) VIA A. BIANCHI, 9 - 25124 BRESCIA Reparto di Virologia e Sierologia Specializzata Tel 030/2290289 - Fax 030/2290535 Email : virol@bs.izs.itvirol@bs.izs.it