Immunodiagnostic monoclonal antibody-based sandwich ELISA of fasciolosis by detection of Fasciola gigantica circulating fatty acid binding protein.
Up to now, parasitological diagnosis of fasciolosis is often unreliable and possesses low sensitivity. Hence, the detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden. In the present study, a monoclonal antibody (MoAb) against recombinant Fasciola gigantica fatty acid binding protein (rFgFABP) has been produced. As well, a reliable sandwich enzyme-linked immunosorbent assay (sandwich ELISA) has been developed for the detection of circulating FABP in the sera of mice experimentally and cattle naturally infected with F. gigantica. MoAb 3A3 and biotinylated rabbit anti-recombinant FABP antibody were selected due to their high reactivities and specificities. The lower detection limit of sandwich ELISA was 5 pg mL-1, and no cross-reaction with other parasite antigens was observed. This assay could detect F. gigantica infection from day 1 post infection. In experimental mice, the sensitivity, specificity and accuracy of this assay were 93·3, 100 and 98·2%, while in natural cattle they were 96·7, 100 and 99·1%. Hence, this sandwich ELISA method showed high efficiencies and precisions for diagnosis of fasciolosis by F. gigantica.
Division of Agricultural Science,Mahidol University,Kanchanaburi Campus,Saiyok,Kanchanaburi 71150,Thailand.
Panat
P
Anuracpreeda
Department of Microbiology,Faculty of Science,Mahidol University,Rama VI Rd.,Bangkok 10400,Thailand.
Runglawan
R
Chawengkirttikul
Department of Anatomy,Faculty of Science, Mahidol University,Rama VI Rd.,Bangkok 10400,Thailand.
Prasert
P
Sobhon
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000911:Antibodies, Monoclonal / Q000276:immunology; D000947:Antigens, Helminth / Q000097:blood / Q000276:immunology; D002417:Cattle / Q000469:parasitology; D002418:Cattle Diseases / Q000175:diagnosis* / Q000276:immunology / Q000469:parasitology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005209:Fasciola / Q000737:chemistry / Q000276:immunology*; D005211:Fascioliasis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D050556:Fatty Acid-Binding Proteins / Q000097:blood* / Q000276:immunology*; D057230:Limit of Detection; D051379:Mice; D012680:Sensitivity and Specificity
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D050556:Fatty Acid-Binding Proteins
Fasciola gigantica; Seroantigen; fatty acid binding protein; immunodiagnosis; monoclonal antibody; sandwich ELISA