title: Production of Monoclonal Antibody Against Excretory-Secretory Antigen of Fasciola hepatica and Evaluation of Its Efficacy in the Diagnosis of Fascioliasis.
abstract: Parasitological methods are not helpful for the diagnosis of fascioliasis in acute and invasive periods of the disease. Detection of coproantigens seems to be a suitable alternative approach in the diagnosis of fascioliasis. The present study aimed to develop a reliable antigen detection system, using monoclonal antibodies raised against excretory-secretory (ES) antigen of Fasciola hepatica, for the diagnosis of fascioliasis. Fasciola adult worms were collected from the bile ducts of infected animals. Species of the fluke was determined by polymerase chain reaction-restriction fragment length polymorphism (RFLP-PCR). ES antigen of F. hepatica was prepared. For production of monoclonal antibodies, mice were immunized with ES antigens of F. hepatica. Spleen cells from the immunized mice were fused with NS-1 myeloma cells, using polyethylene glycol. Hybridoma cells secreting specific antibody were expanded and cloned by limiting dilution. Moreover, polyclonal antibody was produced against F. hepatica ES antigen in rabbits. A capture enzyme-linked immunosorbent assay (ELISA) system, using produced monoclonal antibody, was designed and stool samples of infected animals along with control samples were tested by the system. The capture ELISA detected the coproantigen in 27 of 30 (90%) parasitologically confirmed fascioliasis cases, while 4 of 39 (10.25%) samples infected with other parasitic infections showed a positive reaction in this system. No positive reactivity was found with healthy control samples. Accordingly, sensitivity of 90% and specificity of 94.2% were obtained for the capture ELISA system. The results were compared with those obtained with commercial BIO-X ELISA, and a very good (kappa = 0.9) agreement was found between the commercial kit and the developed capture ELISA. Findings of this study showed that the produced monoclonal antibody has appropriate performance for the detection of Fasciola coproantigen in stool samples and can be appropriately used for the diagnosis of fascioliasis.
authors.affiliation: 1 Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences , Shiraz, Iran .
authors.forename: Samaneh
authors.initials: S
authors.lastname: Abdolahi Khabisi
authors.affiliation: 1 Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences , Shiraz, Iran .|2 Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences , Shiraz, Iran .
authors.forename: Bahador
authors.initials: B
authors.lastname: Sarkari
authors.affiliation: 3 Cellular and Molecular Research Center, Yasuj University of Medical Sciences , Yasuj, Iran .
authors.forename: Abdolali
authors.initials: A
authors.lastname: Moshfe
authors.affiliation: 4 Department of Biotechnology, School of Paramedical Sciences, Shiraz University of Medical Sciences , Shiraz, Iran .
authors.forename: Sedigheh
authors.initials: S
authors.lastname: Jalali
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000947:Antigens, Helminth / Q000276:immunology*; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005210:Fasciola hepatica / Q000276:immunology*; D005211:Fascioliasis / Q000175:diagnosis* / Q000469:parasitology; D005260:Female; D015801:Helminth Proteins / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D012680:Sensitivity and Specificity; D012756:Sheep
chemical_list: D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D015801:Helminth Proteins
keywords: Fasciola; capture ELISA; coproantigen; diagnosis; monoclonal antibody