Enzyme linked immunoassay and fluorescent antibody techniques in the diagnosis of viral diseases using staphylococcal protein-A instead of anti-gamma-globulins.
Staphylococcal protein-A (SpA) is known to interact with the crystallizable fragment (Fc) of IgG molecules from several species. In the present study, SpA coupled to either fluorescein isothiocyanate (FITC) or peroxidase was used in place of antisera to IgG for the fluorescent antibody (FA) techniques and the enzyme linked immunoassay (ELISA). The SpA conjugates produced low background staining when applied in these techniques, and provide a rapid, highly specific and sensitive means for the identification of viral isolates and the detection of serum antibodies. Moreover, SpA is a single reagent that replaces various preparations of anti-gamma globulin against many species. SpA-FITC conjugate was successfully applied for the identification of pseudorabies virus, hog cholera virus, swine vesicular disease virus, transmissible gastroenteritis virus, porcine parvovirus and porcine enteroviruses. Antibody titers against the mentioned viruses could be determined semi-quantitatively in the indirect FA test with SpA-FITC. In our laboratory the ELISA became a routinely practicable serological test for the detection of antibodies only after we introduced SpA-peroxidase as a marker for the IgG.
Bommeli
W
W
Eidgenössisches Vakzine-Institut, CH-4025 Basel, Switzerland.
Kihm
U
U
Zindel
F
F
Fey
H
H
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000918:Antibody Specificity; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000821:virology*; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D005455:Fluorescent Antibody Technique / Q000379:methods / Q000662:veterinary*; D006801:Humans; D013205:Staphylococcal Protein A / Q000276:immunology*; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology*
D000914:Antibodies, Viral; D013205:Staphylococcal Protein A