Precipitation of Radiolabeled Poliovirus with Specific Antibody and Antiglobulin
| Autor: | Robert K. Gerloff, Bill H. Hoyer, Leroy C. McLaren |
|---|---|
| Rok vydání: | 1962 |
| Předmět: | |
| Zdroj: | The Journal of Immunology. 89:559-570 |
| ISSN: | 1550-6606 0022-1767 |
| Popis: | Summary A highly sensitive and specific serologic test for type 2 poliovirus or its antibody is described. Radiolabeled virus in submicrogram quantities was incubated with specific antiserum; thereafter, rabbit antiglobulin was added and the incubation repeated. Coupling between antiglobulin and globulin resulted in precipitation of both, along with virus in combination with immune globulin. Precipitation of radioactivity was used as the measure of virus-antibody reaction, since it paralleled precipitation of infectious particles. Thus the test has been called the radioisotope precipitation (RIP) test. Sensitivity of the test varied inversely with concentration of virus and could be increased to about 80 times that of the tissue culture neutralization test. Ratio of antiglobulin to total globulin was unimportant if antiglobulin were in excess; likewise, quantity of precipitate formed was inconsequential if all of the specific immune globulin were included in it. The test could be completed in 2 to 3 hr, if necessary, and was performed satisfactorily with virus inactivated by carefully controlled ultraviolet irradiation. Inhibition of reaction between labeled virus and its antibody by prior exposure of the antibody to unlabeled virus resulted in conditions whereby unlabeled virus in 0.001 to 0.1 µg quantities could be assayed. Although most of the studies were done with P32-labeled virus, the feasibility and advantages of using I131 for labeling were investigated. Removal of infectious units from the supernatant was also a satisfactory index of presence of specific antibody and enables the use of minute quantities of nonradioactive, biologically active antigen in this type of test. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|