Multispecies detection of antibodies to influenza A viruses by a double-antigen sandwich ELISA

作者：Watcharatanyatip Kamolwan; Boonmoh Sirikwan; Chaichoun Kridsada; Songserm Taweesak; Woratanti Mingkhwan; Dharakul Tararaj^*

来源：Journal of Virological Methods, 2010, 163(2): 238-243.

DOI：10.1016/j.jviromet.2009.09.027

• Influenza virus
• Double-antigen sandwich ELISA
• Antibody detection
• Nucleoprotein

A double-antigen sandwich ELISA was developed for the detection of antibodies to influenza A viruses. A recombinant nucleoprotein (rNP) of influenza A virus was used as a capture antigen and an HRP-conjugate for detecting the antibodies. A total of 125 serum samples from birds of different species including chickens, geese. open-billed storks, Khaki Campbell ducks, lesser whistling ducks, and pigeons with known antibodies were tested by ELISA. The sensitivity and the specificity of ELISA were found to be 98% and 97.3%, respectively. The assay was able to detect the presence of influenza A antibodies as early as the fourth day post-inoculation in ducks infected experimentally with influenza A (H5N1) virus. Excellent agreement (97.6%) was obtained between this sandwich ELISA and the hemagglutination inhibition (HI) tests (kappa = 0.95). The double-antigen sandwich ELISA correlated well with a commercial avian influenza (AI) multispecies ELISA and was slightly more sensitive than the AI multispecies ELISA. These findings indicate that the double-antigen sandwich ELISA based on rNP may offer an effective screening method for serodiagnosis of influenza A virus. The double-antigen sandwich ELISA also enables the detection of antibodies to influenza A viruses in different species without the need for species-specific secondary antibodies.

• 出版日期2010-2