Determination of scrapie agent titer from incubation period measurements in hamsters

Progress in studies on the scrapie agent has been hampered by the slow and tedious endpoint titration assays in rodents. A new assay based on incubation period measurements has been developed. The incubation period is defined as the time interval from inoculation to the onset of clinically detectable neurological illness or as the interval from inoculation to death. Both of these intervals or incubation periods are inversely proportional to the size of the dose injected intracerebrally into random bred weanling syrian hamsters. The incubation period was found to be a linear function of the logarithm of the inoculum size over a wide range of dosages (10(2) - 10(8) ID50 units). From these studies an equation relating the titer of the inoculum to the dilution of the sample and the length of the incubation period has been developed facilitating the use of a computerized record system. Validation of the assay was provided by comparing samples where the agent was measured both by endpoint titration and incubation period methods. Agreement between the 2 methods was generally within +/- 0.5 log10 ID50 units. In addition, no differences between the molecular properties of the agents from hamster and murine sources have been detected using primarily the incubation period method with the former and endpoint titration with the latter. The advantages of the incubation period assay are considerable with respect to time and economy.