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Avian infectious bronchitis (IB) is an acute and highly contagious disease caused by infectious bronchitis virus (IBV). In the study, according to IBV gene sequences published in GenBank, specific primers were designed to clone N gene by RT-PCR, and this gene was inserted into pET-30a (+) vector resulting in a prokaryotic expression plasma pET-30a-N. The results of SDS-PAGE and West Blot analysis showed that the recombinant protein was expressed successfully and had good reactivity with IBV positive serum. Using purified recombinant N protein as a coating antigen, the indirect ELISA protocol was established and optimized, in which N protein was 2.5 μg ? mL-1 of concentration, sample serum of 1 : 40 dilution. For clinical specimen, the IBV antibodies could be detected by this method efficiently and got nearly the same results as those of IBV-mediated ELISA. It would provide a good tool for rapid diagnosis and epidemiological study of avian infectious bronchitis.