title: A 2.9 kDa Fasciola hepatica-recombinant protein based ELISA test for the detection of current-ovine fasciolosis trickle infected.
abstract: The suitability of an enzyme linked immunosorbent assay (ELISA) test with a 2.9 kDa Fasciola hepatica-recombinant protein (FhrAPS) for diagnosing early and current-ovine fasciolosis was analyzed, and compared to that obtained by using a direct ELISA for detecting F. hepatica-circulating FhES antigens and to the coprological sedimentation for fluke egg quantitation. Fourteen Gallega autochthonous breed sheep were experimentally infected with metacercariae by a trickle system (small repetitive infections) and divided into two groups: G-I represented a primary infection for 34 weeks; G-R, animals with primary infection and reinfected 18 w.a.p.i. Seven sheep were left uninfected as the control group (G-C). Serum IgG antibody values against the FhrAPS rose rapidly by 1st w.a.p.i. in all infected sheep. Antibody levels in those with primary infection (G-I, G-C) peaked at 10 weeks, diminishing slightly and levelling from 16 to 34 weeks. Those with primary infection reinfected at 18 weeks had a rebound effect with the highest values observed. Circulating F. hepatica-ES antigens were detected by the 1st w.a.p.i. in all infected groups peaking at 6 weeks, decreasing rapidly to uninfected control values by 10 weeks of infection. Faecal egg-output started 11 weeks after primary infection. An increase in the IgG antibody as well as antigen responses to the FhrAPS and to anti-FhES from the 18 w.a.p.i. was recorded in G-T and G-R after the challenge infection. Antibody levels remained high whereas antigenemia values diminished after 6 weeks. A positive significant correlation between the IgG response against the FhrAPS and the F. hepatica circulating antigens (r2 = 0.428, p = 0.001) was obtained. In conclusion, our standardized diagnostic ELISA for fasciolosis based on the detection of IgG responses to the FhrAPS would be a valuable tool to diagnosis early and current F. hepatica-infections in sheep.
authors.affiliation: Animal Pathology Department, Parasitic diseases, Epidemiology and Zoonoses, Veterinary Faculty, Santiago de Compostela University, 27071-Lugo, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Arias
authors.forename: G V
authors.initials: GV
authors.lastname: Hillyer
authors.forename: R
authors.initials: R
authors.lastname: Sánchez-Andrade
authors.forename: J L
authors.initials: JL
authors.lastname: Suárez
authors.forename: J
authors.initials: J
authors.lastname: Pedreira
authors.forename: C
authors.initials: C
authors.lastname: Lomba
authors.forename: P
authors.initials: P
authors.lastname: Díaz
authors.forename: P
authors.initials: P
authors.lastname: Morrondo
authors.forename: P
authors.initials: P
authors.lastname: Díez-Baños
authors.forename: A
authors.initials: A
authors.lastname: Paz-Silva
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005210:Fasciola hepatica / Q000276:immunology*; D005211:Fascioliasis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D005243:Feces / Q000469:parasitology; D015801:Helminth Proteins / Q000276:immunology*; D010270:Parasite Egg Count / Q000662:veterinary; D011897:Random Allocation; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis* / Q000469:parasitology; D013997:Time Factors
chemical_list: D000947:Antigens, Helminth; D015801:Helminth Proteins; D011994:Recombinant Proteins