Evaluation of a computer-assisted, kinetics-based enzyme-linked immunosorbent assay for detection of coronavirus antibodies in cats
| Autor: | F W Scott, T J Lynch, J E Barlough, D R Downing, R H Jacobson, K L Marcella |
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| Rok vydání: | 1983 |
| Předmět: |
Microbiology (medical)
Feline coronavirus Coronaviridae Infections Color Centrifugation Enzyme-Linked Immunosorbent Assay Antibodies Viral Cat Diseases medicine.disease_cause Serology Antigen-Antibody Reactions Tissue culture Antigen Culture Techniques medicine Animals Bovine serum albumin Antigens Viral Cells Cultured Coronavirus biology Computers Gamma globulin Molecular biology Kinetics Cats biology.protein Antibody Research Article |
| Zdroj: | Journal of Clinical Microbiology. 17:202-217 |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.17.2.202-217.1983 |
| Popis: | A computer-assisted, kinetics-based enzyme-linked immunosorbent assay was adapted for the detection of coronavirus antibodies in feline serum. An alkaline antigen diluent (carbonate-bicarbonate buffer, pH 9.6) used in initial experiments produced diffuse, nonspecific color reactions in both viral and control antigen cuvettes which were correlated, paradoxically, with coronavirus antibody levels in test sera. These interfering reactions were minimized by use of lower-pH antigen diluents such as water and phosphate-buffered saline. Background kinetics-based enzyme-linked immunosorbent assay reactivity directed against a noncoronaviral component of antigen tissue culture fluids could then detected in numerous sera, particularly in samples with lower titers. Much of this reactivity was shown to be associated with bovine gamma globulins in cell culture fluid. It was not serum lot or species specific, since a variety of bovine serum lots as well as individual lots of serum from other mammalian and avian species reacted. Reactivity was markedly reduced when cells for antigen preparation were grown in gamma globulin-free bovine serum. Generation of corrected slope values from the kinetics-based enzyme-linked immunosorbent assay made it possible to correct for residual background reactivity in individual test sera and thus eliminate a potentially major source of false-positive reactions. Collectively, these studies indicated that the control of nonspecific reactivity in feline coronavirus serology is absolutely essential to obtain useful estimates of specific antibody responses. |
| Databáze: | OpenAIRE |
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