In order to obtain pure Eimeria strain,three Eimeria species were collected from the poultry farms infected with coccidia in Chaohu,Huoqiu and Dingyuan of Anhui Province,and then cultured to sporulated oocysts.The 1-7 days old chicks were vaccinated the single-oocyst by single-oocyst separation technology.The oocysts were collected after 7-12 days,and then the coccidia oocysts were identified with biological characteristics and PCR method.The results showed that the ratio of infection were 50%,57% and 29%,respectively in three areas.The three Eimeria species were all identified as Eimeria tenella by biological characteristics and PCR.This indicated that the single oocyst isolation technique was simply and successful.Using specific primers of PCR to identify strains could lay foundation for the study of pure local strains.