title: Establishing an indirect sandwich enzyme-linked-immunosorbent-assay (ELISA) for the detection of antibodies against Histomonas meleagridis from experimentally infected specific pathogen-free chickens and turkeys.
abstract: No serological method suitable for large screening of antibodies against Histomonas meleagridis in poultry is available so far, the objective targeted in the present investigation. Consequently, an ELISA was developed as a suitable tool for this purpose. Investigating serum samples from non-infected specific pathogen-free (spf) chickens and commercial turkeys a high-background signal was noticed when ELISA plates were directly coated with purified parasitic cells. This signal was significantly reduced by coating the plates with a polyclonal rabbit antibody, raised against histomonads, prior to the addition of the antigen. Adopting this approach five antigen preparations were compared and a high reproducibility could be demonstrated reflected by a very low coefficient of variation of 5.3% and 1.7% for the chicken and turkey sera, respectively. After this initial development all further experiments were carried out with one set of plates and the same antigen preparation. Investigating chicken sera obtained from birds infected at 14 days of life, OD values above a predetermined cut-off value were observed 2 weeks post-infection and a rise of IgG antibodies was noticed until 6 weeks post-infection, when the experiment was terminated. Non-protected turkeys infected at 6 weeks of age displayed an increasing IgG response until 14 days post-infection, prior to the death of animals due to histomonosis. In comparison, the majority of turkeys vaccinated with attenuated histomonads, obtained through prolonged passaging and challenged 4 weeks later with virulent parasites, displayed a demonstrable antibody response after the challenge only. Antibody titres increased until 4 weeks post-challenge when the birds were killed and the study was terminated. Altogether, the developed indirect sandwich ELISA proved to be a quick and efficient method to detect IgG antibodies against H. meleagridis in sera of experimentally infected chickens and turkeys and will be a helpful tool to obtain more insights into the epidemiology of the parasite and the immune response of its hosts.
authors.affiliation: Clinic for Avian, Reptile and Fish Medicine, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine, Veterinärplatz 1, A-1210 Vienna, Austria.
authors.forename: M
authors.initials: M
authors.lastname: Windisch
authors.forename: M
authors.initials: M
authors.lastname: Hess
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D011201:Poultry Diseases / Q000175:diagnosis / Q000276:immunology / Q000469:parasitology; D011528:Protozoan Infections / Q000175:diagnosis* / Q000276:immunology / Q000469:parasitology; D013047:Specific Pathogen-Free Organisms; D016834:Trichomonadida / Q000276:immunology*; D014422:Turkeys
chemical_list: D000913:Antibodies, Protozoan