A microtiter, solid-phase, indirect radioimmunoassay (RIA) has been developed and evaluated as a method for detecting poliovirus in water samples. Antiserum to poliovirus Type I, LSc2ab, was prepared in rabbits, and high titer, high avidity antiserum to rabbit globulins were radio- actively labeled with 125I by a modification of the Bolton-Hunter and chloramine T methods to increasing specific activities. The immunoreactivity of the labeled antibodies was assessed. After the preparation and standardization of all components of the assay. The optimum assay conditions were determined. These conditions included the method of coating the microtiter wells with antigen, the time and temperature of incubation of each antiserum, the dilution of each antiserum, and means of reducing non-specific binding. The RIA was conducted with varying numbers of viral plaque forming units. In replicate experiments, average binding ratios of 784% were obtained when different numbers of virus were first reacted with antiserum. This technique shows increased sensitivity over the direct method. These results indicate that the use of RIA to detect viruses in water is indeed feasible.
| Identifer | oai:union.ndltd.org:ucf.edu/oai:stars.library.ucf.edu:rtd-1403 |
| Date | 01 January 1979 |
| Creators | Charba, Sheril K. |
| Publisher | University of Central Florida |
| Source Sets | University of Central Florida |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Retrospective Theses and Dissertations |
| Rights | Written permission granted by copyright holder to the University of Central Florida Libraries to digitize and distribute for nonprofit, educational purposes. |
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