Effect of fixation on the detection of transmissible gastroenteritis coronavirus antigens by the fixed-cell immunoperoxidase technique.
The effect of various fixatives and detergents on the in vitro detection of the viral determinants which are expressed in swine testis cells infected with the transmissible gastroenteritis coronavirus (TGEV) was studied using a microwell immunoperoxidase technique. When compared with glutaraldehyde and formaldehyde, 0.1% paraformaldehyde was found to be the fixative of choice for the detection of these determinants on the membranes of infected cells. Among dehydrating fixatives, 80% acetone or a mixture of acetone and ethanol or of acetone, methanol and ethanol were found to be the best fixatives for the detection of these viral determinants which are expressed in infected cells. In the case of acetone, the temperature of fixation and its concentration in the fixative preparation were found to be important. The treatment of 0.05% glutaraldehyde-fixed, infected cells with 0.1% saponin or 0.1% paraformaldehyde-fixed, infected cells with 1%NP-40 led to satisfactory detection of viral determinants. Using Triton X-100 to render cells permeable, the quantities of N and M antigen detected in TGEV-infected cells prefixed with either 0.05% glutaraldehyde or 0.1% paraformaldehyde were equal to those of 80% acetone-fixed, TGEV-infected cells while the quantity of S antigen detected was diminished. The effect of other detergents such as zwittergent, empigen BB, Chaps and N-lauroylsarcosine on the detection of viral determinants was also studied.
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National Institute of Veterinary Research, Department of Virology, Hanoi, Viet Nam.
Bernard
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D000096:Acetone / Q000737:chemistry; D000447:Aldehydes / Q000737:chemistry; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000737:chemistry / Q000276:immunology*; D003902:Detergents; D004306:Dose-Response Relationship, Immunologic; D005404:Fixatives; D007124:Immunoenzyme Techniques; D005760:Transmissible gastroenteritis virus / Q000276:immunology*
D000447:Aldehydes; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D003902:Detergents; D005404:Fixatives; D000096:Acetone