Examination of syphilitic serum or cerebrospinal fluid can be made at any temperature between 23° and 37°C. The velocity of the fixation reaction, including the fixation of complement and subsequent hemolysis, is greater at a higher temperature, the optimum point being 37°C. The maximum reaction is also reached, however, when the mixture of lipoids, syphilitic serum, and complement is allowed to stand for a long enough period at a lower temperature, the minimum thermal point being near 23°C. For the optimum temperature (37°C.) an incubation of 30 minutes is sufficient, while for the minimum temperature (23°C.) 2 hours are necessary. At the temperature of 30°C. the reaction proceeds with moderate velocity and is complete within 60 minutes.

Guinea pig complement gave a sharper reaction with the sera which contained less than one unit of the fixing substance. Fixation is complete, however, at any of the three temperatures within 20 minutes when there are more than two units present. A serum containing one unit of fixing substance will complete reaction within 30 minutes at 37°C., 60 minutes at 30°C., and 2 hours at 23°C., irrespective of whether human or guinea pig complement is used.

For many reasons a properly adjusted thermostat for 37°C. is recommended for conducting the serum diagnosis of syphilis when possible, but it should not be overlooked that at a temperature near 30°C. an entirely reliable result can be obtained without a special incubator. Even at a temperature as low as 23°C. the test can be carried out if sufficient length of time is allowed.

The foregoing conclusions refer only to the systems in which the acetone-insoluble fraction of tissue lipoids is used as antigen.

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