Isolation of peplomer glycoprotein E2 of transmissible gastroenteritis virus and application in enzyme-linked immunosorbent assay.
An enzyme-linked immunosorbent assay (ELISA) based on peplomer glycoprotein E2 was developed for the detection of antibodies to transmissible gastroenteritis virus (TGEV). Purified preparations of E2 were isolated by solubilization of the viral membrane with nonion detergent Nonidet P-40, followed by sucrose density gradient sedimentation. ELISA optical density values with E2 antigen significantly exceeded the indices when other TGEV protein or intact virion antigen was used. It was shown that a virus protein concentration in the E2 preparation of 500 ng per well is sufficient to sensitize the solid phase of microplates. A comparison of the ELISA and the virus neutralization test for the detection of TGEV antibodies was conducted. A significant correlation between the ELISA and the virus neutralization test was shown (r = 0.97). This serological test may be successfully used for various immunologic investigations.
Rukhadze
G G
GG
All-Union Research Institute of Experimental Veterinary Medicine VASKhNIL, Kuzminki, Moscow, USSR.
Aliper
T I
TI
Sergeev
V A
VA
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D000956:Antigens, Viral / Q000276:immunology; D002499:Centrifugation, Density Gradient; D003332:Coronaviridae / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D008854:Microscopy, Electron; D009500:Neutralization Tests; D005760:Transmissible gastroenteritis virus / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000276:immunology* / Q000302:isolation & purification / Q000648:ultrastructure
D000914:Antibodies, Viral; D000956:Antigens, Viral; D014759:Viral Envelope Proteins