Comparison of serological methods for detection of avian influenza virus antibodies

가금인플루엔자 바이러스 항체검출을 위한 혈청학적 진단법 비교

  • Han, Myung-guk (Avian Disease Division, National Veterinary Research and Quarantine Service) ;
  • Park, Kyoung-yoon (Animal Health Division, Bayer Korea Ltd.) ;
  • Kwon, Yong-kuk (Avian Disease Division, National Veterinary Research and Quarantine Service) ;
  • Kim, Jae-hong (Avian Disease Division, National Veterinary Research and Quarantine Service)
  • 한명국 (국립수의과학검역원 조류질병과) ;
  • 박경윤 (바이엘코리아(주) 임상병리실) ;
  • 권용국 (국립수의과학검역원 조류질병과) ;
  • 김재홍 (국립수의과학검역원 조류질병과)
  • Accepted : 2002.02.27
  • Published : 2002.03.25


An enzyme-linked immunosorbent assay (ELISA) using purified hemagglutinin of swine influenza virus (H1N1) as antigen was developed for detection of antibody to avian influenza virus (AIV). The sensitivity and specificity of a developed and commercial available ELISA kits were compared with those of agar gel precipitation (AGP) test and hemagglutination inhibition (HI) test using sera collected from chickens under condition of field exposure. The concentration of antigen, serum dilution and concentration of enzyme-conjugated secondary antibody in developed ELISA (S-ELISA) were 0.5ug/100ul, 1:200 and 0.03ug/100ul, respectively. The correlation coefficients between S-ELISA and commercial ELISA and HI titers were 0.419 and 0.533, respectively. A significant correlation (p < 0.01) was not found between HI and ELISA titers. The S-ELISA was found to be as more sensitive and specific than the AGP test, showing 86.8% sensitivity and 85.3% specificity. It is suggested that the ELISA using the SIV as antigen may be useful method as an investigating tool for AIV serological surveillance.


  1. Abraham A, Sivanandan V, Halvorson DA, et al. Standardization of enzyme-linked immunosorbent assay for avian influenza virus antibodies in turkey. Am J Vet Res, 47:561-566, 1986.
  2. Abraham A, Sivanandan V, Karunakaran D, et al. Comparative serological evaluation of avian influenza vaccine in turkeys. Avian Dis, 32:659-662, 1988
  3. Abraham A, Sivanandan V, Newman JA, et al. Rapid purification of avian influenza virus for use in enzyme-linked immunosorbant assay. Am J Vet Res,45:959-962, 1984
  4. Alexander DJ. The epidemiology and control of avian influenza and Newcastle disease. J Comp Path, 112:105-126, 1995
  5. Alexander DJ, Allan WH. Avian influenza in turkeys : a survey of farms in eastern England 1979/80. Br Vet J, 138:473-479, 1982
  6. Arenas A, Carranza J, Perea A, et al. Type A influenza viruses in birds in southern Spain: Serological survey by enzyme-linked immunosorbent assay and haemagglutination inhibition tests. Avian Pathol, 19:539-546, 1990
  7. Austin PJ, Webster RG. Antigenic mapping of an avian H1 influenza virus haemagglutination and interrelationship of H1 viruses from Human, Pigs and Birds. J Gen Virol. 67:932-99, 1986
  8. Bean WJ, Kawaoka Y, Wood JM, et al. Characterization of virulent and avirulent A/chicken/Pennsylvania/83 influenza A viruses : potential role of defective interfering RNAs in nature. J Virol, 54:151-160, 1985
  9. Beard CW. Avian influenza antibody detection by immunodiffusion. Avian Dis, 14:337-341, 1970
  10. Beck JR, Swayne DE. Evaluation of ELISA for avian influenza serologic and diagnostic programs: Comparison with agar gel precipitin and hemagglutination inhibition tests. in Proceedings. 4th International Symposium on Avian Influenza, 297-303, 1997
  11. Brown J, Resurreccion RS, Dickson TG. The relationship between the hemagglutination-inhibition test and the enzyme-linked immunosorbent assay for the detection of antibody to Newcastle disease. Avian Dis, 34:585-587, 1990
  12. Easterday BC, Hinshaw SV, Halvorson DA. Influenza. In Calnek BW, Barnes HJ, Beard CW, et al., ed. Diseases of Poultry, 10th ed, Iowa State University Press, Ames, Iowa, 583-605, 1997
  13. Fatunmbi OO, Newman JA, Sivanandan V, et al. A broad- spectrum avian influenza subtype antigen for indirect enzyme-linked immunosorbent assay. Avian Dis, 33:264-269, 1989
  14. Hammond GW, Smith SJ, Noble GR. Sensitivity and specificity of enzyme immunoassays for serodiagnosis of influenza A virus infections. J Infect Dis, 141:644-651, 1980
  15. Kawaoka Y, Nawevw CW, Webster RG. Is virulence of H5N2 influenza viruses in chickens associated with loss of carbohydrate from the hemagglutinin? Virology,139:303-316, 1984
  16. King DJ. Evaluation of different methods of inactivation of Newcastle disease virus and avian influenza virus in egg fluids and serum. Avian Dis, 35:505-514, 1991
  17. Masihi KN, Lange W. Enzyme-linked immunosorbant assay for the detection of influenza type specific antibodies. J Immunol Methods, 36:173-179, 1980
  18. Meulenmans G, Carlier MC, Gonze M, et al. Comparison of hemagglutination-inhibition, agar gel precipitin and enzyme- linked immunosorbent assay for measuring antibodies against influenza viruses in chickens. Avian Dis, 31:560-563, 1987
  19. Mo IP, Song CS, Kim KS, et al. An occurrence of non- highly pathogenic avian influenza in Korea. in Proceedings. 4th International Symposium on AvianInfluenza, 379-383, 1997
  20. Nettles VF, Wood JM, Webster RG. Wildlife surveillance associated with an outbreak of lethal H5N2 avian influenza in domestic poultry. Avian Dis, 29:733-741, 1985
  21. Palese P, Young JF. Variation of influenza A, B and C viruses. Science, 215:1468-1474, 1982
  22. Rohm C, Zhou N, Suss J, et al. Characterization of a novel influenza hamegglutinin, H15 : criteria for determination of influenza A subtypes. Virology, 217:508-516, 1996
  23. Schild GC, Oxford JS, Newman RW. Evidence for antigenic variation in influenza A nucleoprotein. Virology, 93:569-573, 1981
  24. Slemons RD, Easterday BC. Host response differences among five avian species to an influenza virus A/turkey/ Ontario/7732/66(Hav5 N?). Bull WHO,17:521-525, 1972
  25. Synder DB, Marquardt WW, Yancey FS, et al. An enzyme-linked immunosorbent assay for the detection of antibody against avian influenza virus. Avian Dis,29:136-144, 1985
  26. Synder DW, Marquardt WW, Mallinson ET, et al. Rapid serological profiling by enzyme-linked immunosorbent assay. 1. Measurement of antibody activitytiter against Newcastle disease virus in a single serum dilution. Avian Dis, 27:161-170, 1983
  27. Thayer SG, Beard CW. Serologic procedures. In Swayne DE, Glisson JR, Jackwood MW, et al., ed. A laboratory manual for the isolation and identification of avian pathogens, 4th ed, Rose Printing, Tallahassee, Florida, 255-266, 1998
  28. de Wit JJ, Heijmans JF, Mekkes DR, et al. Validation of five commercially available ELISA for the detection of antibodies aganist infectious bursal disease virus (serotype 1). Avian Pathol, 30:543-549, 2001
  29. 윤용덕, 강영배, 김종염 등, 외래동물질병도감. 동양문화인쇄, 서울:70-76,1998