Chicken’s erythrocytes are most abundant cells circulating in the body which participate in inflammation and immune responses. However, expression levels of prostaglandins-related genes in H9N2 infected chicken’s erythrocytes have not been studied yet. So, the present study was designed to investigate the PG-related genes in erythrocytes of H9N2 infected chickens. In current study, qRT-PCR and Western blot analysis were performed to identify the expression levels of PG-related genes (COX-2, GSTA3, PTGDS, PTGES, PTGER3 and PTGER4) in erythrocytes of H9N2 infected chickens at time intervals of 0, 2, 6 and 10 h and P < 0.05 was considered as statistically significant. Expression levels were increased significantly in case of COX-2 at 0 and 6 h, GSTA3 and PTGDS at 0 h and PTGER4 at 2 h of infection, while decreased expression was observed in case of COX-2 at 2 h, GSTA3 at 10 h, PTGDS at 6 and 10 h, PTGES at 2 and 10 h, PTGER3 at 0, 2, 6 and 10 h and PTGER4 at 6 and 10 h of infection. Besides, western blot analysis of COX-2 protein expression further supported the mRNA results. Moreover, this was first study indicating the expression levels of PG-related genes in the H9N2 infected chicken’s erythrocytes. This study may provide a new biomarker to detect H9N2 in chickens. Future studies are in process to know the morphological features, and to evaluate the mechanism exhibited by erythrocytes in response to H9N2 with new experimental evidence in chickens.