An antigen sandwiched ELISA for detection of the total antibodies to HCV was established using genetically engineered proteins and synthesized peptides located in the structural and nonstructural regions of HCV genomes as coated antigens and were labeled by horseradish peroxidase, respectively. A comparison of antigen sandwiched ELISA with indirect ELISA showed that the sensitivity of sandwiched system (1:128) was little higher than that of indirect ELISA (1:64), the specificities of both systems were all 100%. This reagent may detect all classes of antibody to HCV in serum, mainly IgG and IgM and is a new diagnostic reagent for detection and early diagnosis of HCV infection.