Partial characterization of the principal soluble antigens associated with the coronavirus of transmissible gastroenteritis by complement fixation and immunodiffusion

Abstract

A microtiter complement fixation (CF) test to detect transmissible gastroenteritis (TGE) viral antigen was developed, using TGE hyperimmune pig serum as an antibody source. Sera from TGE convalescent pigs did not fix complement by this test. Maximal virus and soluble antigen (SA) titers were obtained 36 to 48 h after inoculation of swine testes cells. Cell-associated virus and SA titers were higher than those in the culture fluid, which had to be concentrated 20x before use as antigen in agar immunodiffusion tests (ID). By sucrose density-gradient centrifugation, the SA had a buoyant density of 1.10 g/ml and could be separated from the virus that banded in the 1.19-g/ml region. Virus and SA from three different isolates of TGE had the same buoyant densities. Heating and proteolytic enzyme digestion established the protein nature of the SA. As assayed by CF and ID, there were stability differences between crude and purified preparations of SA. Antibody prepared in rabbits against the SA neutralized the TGE virus.

Physicochemical properties of transmissible gastroenteritis virus hemagglutinin

“…Second, both infectivity and HA activity had a buoyant density of 1.19 g/ml in a linea sucrose gradient. A virion density of 1.19 g/ml is in agreement with the previous results obtained with Minnesota and Illinois strains of TGE virus [26]. Several coronaviruses, HCV, HEV, BCV, MECV, and IBV, have been demonstrated to possess a virion associated hemagglutinin [3,7,12,20,24,27].…”

“…Three strains of TGE virus were used: the TO strain [4] isolated in Japan, and the Miller [26], and Purdue strains [14] isolated in the U.S.A. All were supplied by Dr. S. Furuuchi, National Institute of Animal Health, Tsukuba, Japan. The TO strain was used in this study, unless otherwise stated.…”

Transmissible gastroenteritis coronavirus : the development and applications of the fixed-cell immunoperoxidase technique

“…The literature concerning the detection of anti-TGEV antibodies in swine serum includes several immunological methods such as SN using cell culture-adapted viruses by a variety of procedures (75,16,27,10,11,69), indirect IF (4), bentonite agglutination (59), passive haemagglutination (34), the indirect micro-haemagglutination test (57), complement fixation (62), leukocyte aggregation (73) and agar immunodiffusion (9). Obviously, the application of these techniques in the detection of anti-TGEV antibodies in a great number of samples (e.g.…”

Evaluation of an enzyme-linked immunosorbent assay for the detection of transmissible gastroenteritis virus antibodies

“…Various serological tests for the detection of TGE virus antibodies are available. They include the neutralization test (Harada et al, 1967;Bohl et al, 1972), bentonite agglutination test (Sibinovic et al, 1966), complement fixation test (Stone et al, 1976), agar immunodiffusion test (Bohac et al, 1975;Bohac and Derbyshire, 1976;Stone et al, 1976), indirect fluorescent antibody test (Benfield et al, 1978), indirect haemagglutination test (Shimizu and Shimizu, 1977), and immunoperoxidase test (Kodama et al, 1980). The enzyme-linked immunosorbent assay (ELISA) has been shown to be a useful serological tool for viral infections.…”