J Vet Sci.  2009 Dec;10(4):323-329. 10.4142/jvs.2009.10.4.323.

Evaluation of a competitive ELISA for antibody detection against avian influenza virus

• Song DS
• Lee YJ
• Jeong OM
• Kim YJ
• Park CH
• Yoo JE
• Jeon WJ
• Kwon JH
• Ha GW
• Kang BK
• Lee CS
• Kim HK
• Jung BY
• Kim JH
• Oh JS

• KMID: 1726905
• DOI: http://doi.org/10.4142/jvs.2009.10.4.323

Abstract

Active serologic surveillance is necessary to control the spread of the avian influenza virus (AIV). In this study, we evaluated a commercially-available cELISA in terms of its ability to detect AIV antibodies in the sera of 3,358 animals from twelve species. cELISA detected antibodies against reference H1- through H15-subtype AIV strains without cross reactivity. Furthermore, the cELISA was able to detect antibodies produced following a challenge of the AIV H9N2 subtype in chickens, or following vaccination of the AIV H9 or H5 subtypes in chickens, ducks and geese. Next, we tested the sensitivity and specificity of the cELISA with sera from twelve different animal species, and compared these results with those obtained by the hemagglutination-inhibition (HI) test, the "gold standard" in AIV sera surveillance, a second commercially-available cELISA (IZS ELISA), or the agar gel precipitation (AGP) test. Compared with the HI test, the sensitivities and specificities of cELISA were 95% and 96% in chicken, 86% and 88% in duck, 97% and 100% in turkey, 100% and 87% in goose, and 91% and 97% in swine, respectively. The sensitivities and specificities of the cELISA in this study were higher than those of IZS ELISA for the duck, turkey, goose, and grey partridge sera samples. The results of AGP test against duck and turkey sera also showed significant correlation with the results of cELISA (R-value >0.9). In terms of flock sensitivity, the cELISA correlated better with the HI test than with commercially-available indirect ELISAs, with 100% flock sensitivity.

Keyword

avian influenza virus; competitive ELISA; surveillance

MeSH Terms

Animals
Antibodies, Viral/*blood
Birds
Enzyme-Linked Immunosorbent Assay/methods/*veterinary
Horses
Influenza A virus/*immunology
Influenza Vaccines/immunology
Influenza in Birds/blood/*immunology/prevention & control
Sensitivity and Specificity
Serologic Tests
Species Specificity
Swine

Figure

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  Fig. 1 Seroconversion tests after avian influenza virus (AIV) H9N2 challenge in chickens. The percent inhibition (PI) value was determined by cELISA, and a PI value greater than 50 was regarded as positive. Seroconversion was detected in 2/5 chickens 4 days post-challenge when tested with cELISA and 1/5 chickens when tested by the hemagglutination-inhibition (HI) test. The PI value and HI titer were mean titers from the cELISA and HI test, respectively.

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