Detection of NP, N3 and N7 antibodies to avian influenza virus by indirect ELISA using yeast-expressed antigens Export

@article{citeulike:5915738, abstract = {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.}, author = {Upadhyay, Chitra and Ammayappan, Arun and Vakharia, Vikram}, citeulike-article-id = {5915738}, citeulike-linkout-0 = {http://dx.doi.org/10.1186/1743-422X-6-158}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/19811655}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=19811655}, day = {07}, doi = {10.1186/1743-422X-6-158}, issn = {1743-422X}, journal = {Virology Journal}, keywords = {antibody, influenza, yeast}, month = {October}, number = {1}, pages = {158+}, posted-at = {2009-10-14 09:36:47}, priority = {2}, title = {Detection of NP, N3 and N7 antibodies to avian influenza virus by indirect ELISA using yeast-expressed antigens}, url = {http://dx.doi.org/10.1186/1743-422X-6-158}, volume = {6}, year = {2009} }

TY - JOUR ID - citeulike:5915738 L3 - citeulike-article-id:5915738 N2 - 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. IS - 1 JF - Virology Journal SN - 1743-422X TI - Detection of NP, N3 and N7 antibodies to avian influenza virus by indirect ELISA using yeast-expressed antigens VL - 6 SP - 158 KW - antibody KW - influenza KW - yeast AU - Upadhyay, Chitra AU - Ammayappan, Arun AU - Vakharia, Vikram PY - 2009/10/07/ UR - http://dx.doi.org/10.1186/1743-422X-6-158 ER -