Given that no information on tapeworm DHFR is available, the aims of the present study were (i) to analyse the enzymatic activity characteristics of DHFR from E. granulosus (Eg-DHFR); (ii) to assess the potential of Eg-DHFR as a target for E. granulosus control chemicals; (iii) to express recombinant Eg-DHFR (rEg-DHFR) and locate the native protein in different stages of the parasite; and (iv) to detect the immunogenicity of rEg-DHFR and develop an indirect enzyme-linked immunosorbent assay (ELISA) for diagnosis of CE in sheep. Here, DHFR is linked to cholesteryl ester measurement.