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Background  This research was carried out to develop a reliable monoclonal antibody (MoAb)-based sandwich enzyme linked immunosorbent assay (ELISA) for the diagnosis of active  Fasciola gigantica  infection in both serum and stool for comparative purposes.    Methods  From a panel of MoAbs raised against  F. gigantica  excretory/secretory antigens (ES Ags), a pair (12B/11D/3F and 10A/9D/10G) was chosen due to its high reactivity and strict specificity to  F. gigantica  antigen by indirect ELISA.    Results  The two MoAbs were of the IgG 1  and IgG 2a  subclasses, respectively. Using SDS-PAGE and EITB, the selected MoAbs recognized 83, 64, 45 and 26 kDa bands of ES Ags. The lower detection limit of ELISA assay was 3 ng/ml. In stool, the sensitivity, specificity and diagnostic efficacy of ELISA was 96%, 98.2 and 97.1%; while in serum they were 94%, 94.6% and 94.3%, respectively. Moreover, a positive correlation was found between ova count in stool of  F. gigantica  infected patients and the OD readings of ELISA in both stool and serum samples ( r  = 0.730, p < 0.01 and r = 0.608; p < 0.01, respectively).    Conclusions  These data showed that the use of MoAb-based sandwich ELISA for the detection of  F. gigantica  coproantigens in stool specimens was superior to serum samples; it provides a highly efficient, non-invasive technique for the diagnosis of active  F. gigantica  infection.

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Diagnostic efficacy of monoclonal antibody based sandwich enzyme linked immunosorbent assay (ELISA) for detection of Fasciola gigantica excretory/secretory antigens in both serum and stool