Through the investigations on the diagnostic methods of rabies by complement fixation tests (CFT), especially on the Ando's mehod, it was found that C F-antigenicity decreased by the treatment of the antigen with amyl-alcohol. From this evidence, phospholipid of the antigen was considered to be concerned in the reaction, and the following experiments were conducted.
1) Using the solvents for lipids, such as amyl-alcohol, chloroform, ether, and petroleum ether, the lipid was extracted from the antigens. The effect of the extracted materials on the CFT titers was studied.
2) The analysis of the chemical components of the antigens extracted with those solvents was carried out. The investigations were made on the relationship between the amount of phospholipid, N-contents of proteins, and the C F-antigenicities, and also on the relationship between lipo-proteins and C F-antigenicities.
The results were as follows:
1) A decrease in the antigenicity of C F-antigen of rabies was observed, if the antigen was treated with solvents such as amyl-alcohol, chloroform, ether, and petroleum ether.
2) The amount of phospholipid of the antigen digested with trypsin, and of the putrefied brain antigen was larger than that of the antigen prepared by the Ando's method, also decreased by the treatment with amyl-alcohol.
3) From the result that the amount of phospholipid was in proportion to the CF-antigensicity, the phospholipid was assumed to be the main component of the CFT-antigens.
4) The phospholipid transformed by the rabies virus infection was considered to play an important role in the reaction.
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