2 The Philippine Journal of Science isie 



tube with the blood was left at room temperature for two or 

 three hours to allow the blood to clot. If the clot adhered to 

 the tube and did not sink in the serum, it was loosened with a 

 sterile wire and was pushed down into the serum. The specimen 

 was now put into the refrigerator for from twelve to twenty- 

 four hours to allow the serum to separate and the loose cor- 

 puscles to settle to the bottom of the tube. After the corpuscles 

 had settled, the clear serum was pipetted off and was mixed with 

 the preservative, after which it was tested at suitable intervals. 



Antigen. — The antigen used was plain alcoholic extract of 

 human heart-muscle, and about one fourth the anticomplemen- 

 tary dose was used per tube in the test. 



Complement. — As alien complement the pooled sera of three 

 guinea pigs were used in quantities of 0.1, 0.05, and 0.025 cubic 

 centimeter. 



Haemolytic amboceptor. — As artificial hsemolytic amboceptor 

 the antihuman amboceptor advocated by Noguchi was used in 

 doses of 1 to 1.5 unit per tube. The smallest quantity of am- 

 boceptor that with 0.05 cubic centimeter of complement dis- 

 solved the test dose of corpuscles in one hour was called a unit. 



Corpuscles. — Human corpuscles were well washed, and 0.5 

 cubic centimeter of a 4 per cent suspension in physiologic salt 

 solution was used per tube. 



Glassware. — Two sets of test tubes were used — one set as an- 

 tigen tubes and another set as control tubes. Test tubes that 

 had once been used as antigen tubes were never used as control 

 tubes. Other glassware, such as pipettes, graduates, beakers, 

 and flasks, that was used for serum was used for serum only, 

 and that which was used for antigen was used for antigen only. 



Methods. — Two methods of conducting the serum test for 

 syphilis were used — namely, the Wassermann method with 

 human haemolytic system and the method described by Tschem- 

 ogubow,^ Hecht,^ Gurd,* and others. As human serum as a 

 rule dissolves sheep corpuscles better than the corpuscles of the 

 guinea pig, sheep corpuscles were used, as first advocated by 

 Tschemogubow. 



Technique of conducting the Wassermann reaction. — Unless 

 unheated serum was tested for anticomplementary properties, 

 the serum was heated to between 55° C. and 56° C. for thirty 

 minutes before testing. Six test tubes, three, designated as 

 1, 2, and 3 antigen tubes, and three, designated as 1', 2', and 3' 



'Deut. med. Wochenschr. (1909), 35, 668. 

 'Wien. klin. Wochenschr, (1909), 22, 338. 

 *Joum. Infect. Dis. (1911), 8, 427. 



