Development of Diagnostic Method for Unconventional Agent (Scrapie Gagent) Infection.

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The usefulness of detecting the protein (PrP) consisting of scrapie-associated fibrils in the lymphoreticular organs of sheep as a diagnostic tool was investigated. The PrP was detected by means of a rabbit-anti-sheep PrP polyclonal antibody by Western blot analysis. PrP was detected in samples from the central nervous system of five of six sheep showing clinical signs of natural scrapie infection, in spleen samples from four of the six sheep and in lymph node samples taken from three of the sheep. PrP was detected in the spleen and lymph node samples, but not in the central nervous system samples from one of the six sheep that was clinically and histopathologically abnormal. This animal appeared to be in the early clinical stage of the disease. A total of 47 clinically normal sheep were examined for the presence of PrP. It was detected in spleen samples from three of the 47 sheep and in lymph node samples from three of the 37 sheep tested. Similarly, PrP was detected in a samples of lymph node obtained surgically from one of three experimentally infected sheep 14 months after inoculation. The PrP-positive sheep and one of the remaining PrP-negative sheep showed clinical signs of scrapie six and five months later respectively. One sheep sacrificed 18 months after experimental infection was positive for PrP in the central nervous system, spleen and lymph node, but five other sheep which were sacrificed or died two, eight, 16, 18 and 21 months after infection were negative or doubtful for the detection of PrP.