To establish a real-time fluorescent quantitative PCR assay for detection of IL–1β and IL–8 genes of Cyprinus carpio var. Jian, specific primers were designed according to the gene sequences available in GenBank, and the β–actin gene was used as a reference gene. The standard curve and the melting curve were analyzed through SYBR Green I method. Melting curve showed specific PCR products of β–actin, IL–1β and IL–8 gene were obtained with Tm 86, 82.5 and 84 ℃, respectively. Standard curve there were good linear relationship for β–actin, IL–1β and IL–8 (r2> 0. 990) under detecting Ct range of 12 to 32. The amplification efficiency were 96.3%, 103.2%, and 102.6%, respectively, and the coefficients of variation (CV) of using this developed assay to detect these genes are 0.14%-0.86%, 0.18%-0.93% and 0.13%-0.86%, respectively. Relative expression value of IL–1β and IL–8 in healthy Cyprinus carpio var. Jian was 1.07 and 1.71, respectively. Conclusion: The developed real-time fluorescent quantitative PCR assay for IL–1β and IL–8 of Cyprinus carpio var. Jian, which showed broad range, high efficiency, specificity and reliability, could be used to detect IL–1β and IL–8 expression at mRNA level.