title: Solid phase immune electron microscopy for diagnosis of transmissible gastroenteritis in pigs.
abstract: A serological trapping technique is described for detecting transmissible gastroenteritis (TGE) virus in faeces. The technique involves the coating of electron microscope grids with protein A and specific TGE virus antiserum. Optimal conditions for performing this solid phase immune electron microscopy technique were a concentration 250 micrograms ml-1 of protein A; 1:100 diluted rabbit anti-TGE virus hyperimmune serum for coating the grids and overnight incubation with virus samples. The possibility of detecting coronavirus in crude faeces was highly improved with solid phase immune electron microscopy, compared with conventional negative staining electron microscopy, by specific trapping of virus and prevention of adsorption of contaminants. The sensitivity of the method was evaluated by coded investigation of a dilution series of stock virus mixed with different pools of faeces. The improvement of virus detection in faeces by solid phase immune electron microscopy, compared with standard electron microscopy was at least 100-fold. Faecal shedding of coronavirus by pigs infected with virulent and attenuated strains of TGE virus was studied. Virus detection in faeces by a standard electron microscopy technique was not practical, since the virus was obscured by a large quantity of debris in the faeces. By using solid phase immune electron microscopy, however, the aspect of the specimens on the grids improved so much that, in addition to more common immunofluorescence, the technique might be useful as a diagnostic test for TGE. Virus was detected in daily faecal samples from one or two days after experimental infection with virulent TGE virus, until death in five out of eight animals. Unlike immunofluorescence, solid phase immune electron microscopy may be used for diagnosis in living animals.
authors.affiliation: Central Veterinary Institute, Virology Department, Lelystad, The Netherlands.
authors.forename: A P
authors.initials: AP
authors.lastname: van Nieuwstadt
authors.forename: J B
authors.initials: JB
authors.lastname: Cornelissen
authors.forename: J
authors.initials: J
authors.lastname: Vreeswijk
mesh_terms: D000818:Animals; D005243:Feces / Q000382:microbiology; D005761:Gastroenteritis, Transmissible, of Swine / Q000175:diagnosis*; D008854:Microscopy, Electron / Q000379:methods; D013552:Swine; D005760:Transmissible gastroenteritis virus / Q000276:immunology / Q000648:ultrastructure