Detection of fasciola-specific excretory/secretory [e/s] protein fraction band [49.5 kda] and its utilization in diagnosis of early fascioliasis using different diagnostic techniques

ABSTRACT

The objective of the present work was to evaluate Fasciola E/S antigens for diagnosis of early fascioliasis utilizing different diagnostic techniques. Using enzyme-linked immunoelectrotransfer blot [EITB], Fasciola-specific E/S protein fraction band [49.5 kDa] was determined and electroeluted. The monospecific antibodies against this specific fraction band were prepared by immunizing pathogen-free rabbit. Assessment of the prepared monospecific antibodies in diagnosis of human fascioliasis was performed through the detection of E/S coproantigens by enzyme-linked immunosorbent assay [ELISA] in stool eluates obtained from patients with confirmed fascioliasis, other parasites as well as from other healthy individuals. Serum samples were collected and tested to detect serum antibodies against Fasciola E/S antigen using EITB and counter immunoelectrophoresis [CIEP]. Analysis of Fasciola adult worm E/S products by SDS/PAGE revealed a number of bands, the molecular weight [MW] of which ranged from 14-200 kDa with 3 major bands [27.5, 32.5 and 55 kDa]. Fasciola E/S 49.5 kDa protein fraction proved to be specific of F. gigantica. Cross reaction with S. mansoni was observed at higher MW [110-120 kDa]