Comparison of two methods for detection of transmissible gastroenteritis virus in feces of pigs with experimentally induced infection.
An indirect, double-antibody sandwich-type ELISA for detection of transmissible gastroenteritis virus (TGEV) was developed, using a solid phase of rabbit hyperimmune serum and a pool of 3 antipeplomer monoclonal antibodies to trap and to detect the virus, respectively. The technique was used to detect viral antigen in feces of pigs that had been infected with the virulent Miller strain, the attenuated Purdue strain, or the Erica strain (a Dutch field isolate) of TGEV. The results were compared with those of a solid-phase immunosorbent electron microscopy (SPIEM) technique for virus detection. Both techniques detected shedding of virulent virus in feces obtained from pigs on the first or second day after infection, and virus excretion continued for 6 to 8 consecutive days. Virus shedding started later in pigs infected with the attenuated Purdue strain of TGEV and lasted only 2 to 4 days. In comparison with the 2 virulent strains, infection with the attenuated strain appeared to be limited to a smaller portion of the small intestine. Of 242 fecal specimens that were tested by use of ELISA and SPIEM, 119 had positive results in both tests. Additionally, virus could be detected by ELISA in 21 and by SPIEM in 16 specimens. Fecal specimens obtained from pigs before infection always reacted negatively by ELISA for TGEV antigen; there was no cross-reactivity with fecal specimens containing porcine rotavirus or porcine epidemic diarrhea virus. The ELISA and SPIEM were found to be specific and sensitive for the detection of TGEV in feces.
van Nieuwstadt
A P
AP
Virology Department, Central Veterinary Institute, Lelystad, The Netherlands.
Cornelissen
J B
JB
Zetstra
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000032:analysis*; D003332:Coronaviridae / Q000302:isolation & purification*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005243:Feces / Q000382:microbiology*; D005761:Gastroenteritis, Transmissible, of Swine / Q000175:diagnosis*; D005856:Germ-Free Life; D015151:Immunoblotting; D007163:Immunosorbent Techniques; D008854:Microscopy, Electron; D011237:Predictive Value of Tests; D013047:Specific Pathogen-Free Organisms; D013552:Swine; D005760:Transmissible gastroenteritis virus / Q000276:immunology / Q000302:isolation & purification* / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral