Academic journal article Iranian Journal of Public Health

Development of Enzyme-Linked Immunosorbent Assay with a Nucleoprotein as an Antigen for Detection of Antibodies against Avian Influenza Virus

Academic journal article Iranian Journal of Public Health

Development of Enzyme-Linked Immunosorbent Assay with a Nucleoprotein as an Antigen for Detection of Antibodies against Avian Influenza Virus

Article excerpt

Background: The aim of this study was to produce recom- binant nucleoprotein (NP) and development of an ELISA test with it.

Methods: For this purpose, the coding region of nucleopro- tein (NP) gene of A/chicken/Iran/AH-1/06(H9N2) AI iso- late was amplified by RT-PCR and cloned into a prokaryotic expression vector (pMal-C2x) and transformed into E. coli BL21(DE3). The recombinant protein MBP-NP was ex- pressed in E. coli and was purified using amylase resin chro- matography column and used as the diagnostic antigen to develop a NP-based type specific indirect enzyme linked im- munosorbent assay (ELISA) for detecting antibodies to AI for chicken sera. The NP-ELISA was compared with a commercial AIV ELISA kit (IDEXX- USA).

Results: Excellent agreement was obtained between NP- ELISA and commercial ELISA kit (k= 0. …

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