The elimination of the foreign complement and corpuscles from the test for the serodiagnosis of syphilis has been attempted, and the results so far obtained are very satisfactory. Instead of using guinea pig complement, fresh human serum was utilized for the source of complement for the production of hemolysis upon the human corpuscles in the presence of an adequate amount of the specific anti-human amboceptor (prepared in rabbits). Usually 0.1 cc. of fresh human serum contains enough complement to hemolyze 1 cc. of a 1 per cent suspension of human corpuscles, but the amount of anti-human amboceptor required in this combination is about five to seven times that necessary when guinea pig complement (0.04 cc.) is used. It has also been shown that when a given human serum contains insufficient complement, an adequate quantity (0.1 cc. is usually enough) of another fresh negative serum may be added to supplement it. However, one rarely encounters this group of sera. Inactivated human sera can also be examined by utilizing human complement from another source (the serum must be fresh, active, and negative).
The only drawback to the present method is the comparatively large amount of anti-human hemolytic amboceptor required. It is estimated that 30 to 40 cc. of the anti-human hemolytic immune serum, from one rabbit, of high potency—say 0.005 cc.—would be enough to examine about 3,000 to 4,000 cases (0.01 cc. for each case), whereas if guinea pig complement were used the same amount would cover about 15,000 tests (0.002 cc. for each case). In a large hospital or in the Army there should be no difficulty in preparing any amount of the anti-human hemolytic amboceptor. For example, material for 100,000 tests could be prepared within 1 month in less than 100 rabbits. The amboceptor serum can be used in the fluid state, or, if the titer is high, impregnated into filter papers.
Special attention should be called to the fact that to obtain a powerful anti-human hemolytic amboceptor five to six intraperitoneal injections of corpuscles, thoroughly washed (until there is no trace of serum in the supernatant fluid), in doses of 5, 7, 10, 10, 10, and possibly another 10 cc. of the concentrated suspension (restored to the. original volume of the blood) are required. The bleeding may be done by the 9th or 10th day. The animals may be kept after bleeding for the production of more amboceptor by subsequent injections of the washed corpuscles.
Finally, it should be emphasized that only the acetone-insoluble fraction of tissue lipoids of required standards (Noguchi) should be used when utilizing the human complement in the fixation test.
