Giuseppe
Emeritus
Detection of NP, N3 and N7 antibodies to avian influenza virus by indirect ELISA using yeast-expressed antigens (BMC, Abstract, edited)
Detection of NP, N3 and N7 antibodies to avian influenza virus by indirect ELISA using yeast-expressed antigens
Chitra Upadhyay, Arun Ammayappan and Vikram N Vakharia
Virology Journal 2009, 6:158
doi:10.1186/1743-422X-6-158
Published: 7 October 2009
Abstract (provisional)
Background
Avian influenza viruses, belonging to the family Orthomyxoviridae, possess distinct combinations of hemagglutinin (H) and the neuraminidase (N) surface glycoproteins. Typing of both H and N antigens is essential for the epidemiological and surveillance studies. Therefore, a rapid, sensitive, and specific method for their assay is desirable and an enzyme-linked immunosorbent assay (ELISA) can be useful for this purpose, using recombinant proteins.
Results
The nucleoprotein (NP) and truncated neuraminidase subtype 3 and 7 of avian influenza virus (AIV) were expressed in Saccharomyces cerevisiae (S. cerevisiae) and used to develop an indirect ELISA for antibody detection. The developed assays were evaluated with a panel of 61 chicken serum samples. The performance of NP-ELISA was compared with the commercially available ProFlok(R) AIV ELISA kit. The results showed comparable agreement and sensitivity between the two tests indicating that NP-ELISA assay can be used for screening the influenza type A antibody in AIV-infected birds. The N3- and N7- ELISAs also reacted specifically to their type-specific sera and did not exhibit any cross-reaction with heterologous neuraminidase subtype specific sera.
Conclusion
The study demonstrates the expression of the NP, N3, and N7 proteins of AIV in S. cerevisiae and their application in developing an indirect ELISA for detecting NP, N3 and N7 antibodies from AIV-infected chicken sera. The described indirect ELISAs are rapid, sensitive, specific and can be used as promising tests during serological surveillance.
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<cite cite="http://www.virologyj.com/content/6/1/158">Abstract | Detection of NP, N3 and N7 antibodies to avian influenza virus by indirect ELISA using yeast-expressed antigens</cite>Chitra Upadhyay, Arun Ammayappan and Vikram N Vakharia
Virology Journal 2009, 6:158
doi:10.1186/1743-422X-6-158
Published: 7 October 2009
Abstract (provisional)
Background
Avian influenza viruses, belonging to the family Orthomyxoviridae, possess distinct combinations of hemagglutinin (H) and the neuraminidase (N) surface glycoproteins. Typing of both H and N antigens is essential for the epidemiological and surveillance studies. Therefore, a rapid, sensitive, and specific method for their assay is desirable and an enzyme-linked immunosorbent assay (ELISA) can be useful for this purpose, using recombinant proteins.
Results
The nucleoprotein (NP) and truncated neuraminidase subtype 3 and 7 of avian influenza virus (AIV) were expressed in Saccharomyces cerevisiae (S. cerevisiae) and used to develop an indirect ELISA for antibody detection. The developed assays were evaluated with a panel of 61 chicken serum samples. The performance of NP-ELISA was compared with the commercially available ProFlok(R) AIV ELISA kit. The results showed comparable agreement and sensitivity between the two tests indicating that NP-ELISA assay can be used for screening the influenza type A antibody in AIV-infected birds. The N3- and N7- ELISAs also reacted specifically to their type-specific sera and did not exhibit any cross-reaction with heterologous neuraminidase subtype specific sera.
Conclusion
The study demonstrates the expression of the NP, N3, and N7 proteins of AIV in S. cerevisiae and their application in developing an indirect ELISA for detecting NP, N3 and N7 antibodies from AIV-infected chicken sera. The described indirect ELISAs are rapid, sensitive, specific and can be used as promising tests during serological surveillance.
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