Abstract
A diagnostic immunoaassay using recombinant nucleocapsid protein to detect antibodies to I13V was designed. The nucleocapsid protein was expressed from an expression plasmid, Qiagen pQE8, in E. coli as a fusion protein that was purified on NI-NTA resin. This purified protein was used to determine the optimal conditions for the reagents required in an indirect ELISA and dot blot assay. Based on the optimization of the reagents, a tentative test protocol was derived. 'Me recombinant nucleocapsid protein was found to detect anti-IBV antibodies in polyclonal chicken antisera for various strains of IBV, including Gray, Mass4 1, Conn and Ark99. Negative control serurn with no anti-113V antibodies did not react with the nucleocapsid protein, neither did non-specific proteins like BSA. This study has shown that anti-MV antibodies can be detected with very low concentrations of the recombinant nucleocapsid protein in enzyme immunoassays.
Ndifuna, Abdul-El-Nuru (1996). Serodiagnosis of avian infectious bronchitis virus using recombinant nucleocapsid protein. Master's thesis, Texas A&M University. Available electronically from
https : / /hdl .handle .net /1969 .1 /ETD -TAMU -1996 -THESIS -N38.