Establishement Of A TaqMan Real-time PCR Assay For Detecting Eimeria Stiedai

Rabbit coccidiosis is a disease caused by Eimeria protozoan parasite. Among rabbit coccidian, Eimeria stiedai is the most virulent. Eimeria stiedai parasitizes in the bile duct epithelial cells of rabbits, and may cause serious liver coccidiosis. To improve the the detection efficiency for Eimeria stiedai, we designed the primers and TaqMan probe based on the COI sequence of Eimeria stiedai and established a TaqMan real-time PCR assay for detecting Eimeria stiedai in the study.The result showed that Serial diluted plasmids ranging from 107copies/μL to 10 copies/μL were used as standards. The correlation coefficient was 0.999 and the efficiency of PCR was 88.472%. The standard curve was achieved and the sensitivity of this method was 10 copies/μL, Whilst it was specific for Eimeria stieda and there was no crossing-reaction with other Eimeria spp.. The intra-variation was 0.66%. Pathological examinations revealed hepatomegaly, with presence of discrete yellowish-white nodules of varying size on the surface and throughout the parenchyma.With the time going, lesions were becoming ever more obvious and oocysts were also seen. The results of TaqMan real-time PCR assay were consistent with pathology detections.In general, The TaqMan real-time PCR assay we established for detecting Eimeria Stiedai is a rapid, sensitive and specific method and could be used in prevention and control rabbit coccidiosis induced by Eimeria stiedai.