title: Immunodiagnosis of Fasciola gigantica Infection Using Monoclonal Antibody-Based Sandwich ELISA and Immunochromatographic Assay for Detection of Circulating Cathepsin L1 Protease.
abstract: BACKGROUND
  Tropical fasciolosis caused by Fasciola gigantica infection is one of the major diseases infecting ruminants in the tropical regions of Africa and Asia including Thailand. Parasitological diagnosis of fasciolosis is often unreliable and possesses low sensitivity. Therefore, the detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden.
  
  
  METHODS
  In this study, we have produced a monoclonal antibody (MoAb) against recombinant F. gigantica cathepsin L1 (rFgCatL1), and developed both sandwich enzyme-linked immunosorbent assay (sandwich ELISA) and immunochromatographic (IC) test for rapid detection of circulating cathepsin L1 protease (CatL1) in the sera from mice experimentally and cattle naturally infected with Fasciola gigantica. MoAb 4E3 and biotinylated rabbit anti-recombinant CatL1 antibody were selected due to their high reactivities and specificities.
  
  
  RESULTS
  The lower detection limits of sandwich ELISA and IC test were 3 pg/ml and 0.256 ng/ml, respectively. Sandwich ELISA and IC test could detect F. gigantica infection from day 1 to 35 post infection. In experimental mice, the sensitivity, specificity and accuracy were 95%, 100% and 98.6% (for sandwich ELISA), and 93%, 100% and 98.2% (for IC test), while in natural cattle they were 98.3%, 100% and 99.5% (for sandwich ELISA), and 96.7%, 100% and 99.1% (for IC test).
  
  
  CONCLUSIONS
  These two assay methods showed high efficiencies and precisions for diagnosis of fasciolosis by F. gigantica.
authors.affiliation: Division of Agricultural Science, Mahidol University, Kanchanaburi Campus, Saiyok, Kanchanaburi, Thailand.|Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Bangkok, Thailand.
authors.forename: Panat
authors.initials: P
authors.lastname: Anuracpreeda
authors.affiliation: Department of Microbiology, Faculty of Science, Mahidol University, Rama VI Road, Bangkok, Thailand.
authors.forename: Runglawan
authors.initials: R
authors.lastname: Chawengkirttikul
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Bangkok, Thailand.
authors.forename: Prasert
authors.initials: P
authors.lastname: Sobhon
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D002403:Cathepsins / Q000097:blood / Q000276:immunology*; D002417:Cattle; D002846:Chromatography, Affinity; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005209:Fasciola / Q000201:enzymology / Q000276:immunology* / Q000502:physiology; D005211:Fascioliasis / Q000175:diagnosis / Q000276:immunology* / Q000469:parasitology; D015801:Helminth Proteins / Q000097:blood / Q000276:immunology*; D006790:Host-Parasite Interactions / Q000276:immunology; D008297:Male; D008647:Mesocricetus; D051379:Mice; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D015801:Helminth Proteins; D002403:Cathepsins