208 PUBLIC HEALTH BACTERIOLOGY 



54 C. Serum can also be obtained from cerebrospinal 

 fluid, etc. 

 PROCESS. 



(a). In a test tube, put the following : 



o-i c.c. of complement (fresh guinea-pig serum). 

 0-2 c.c. of inactivated test serum (from patient). 

 i'O c.c. of standardized antigen (liver extract, etc.). 

 Add salt solution to make up bulk to 3 c.c. ; 

 shake thoroughly, and place for I hour in 

 incubator at 37 '5 C. 

 (b). Now add, 



i-o c.c. of 5 per cent emulsion of sheep's red cells. 

 2'0 units of haemolytic serum (determined as 



above) . 

 Shake thoroughly, and incubate again at 37-5 C. 



for i to 2 hours. 

 (c). Observe result. 



(i). No hamolysis. Immune-body or anti- 

 body is present ; test positive. 

 (2). Complete hcemolysis. Immune - body or 



antibody is absent ; test negative. 

 Several controls should be put up at the same time to 

 preclude error. Such are : a test with known normal 

 serum ; tests with antigen and complement alone to see 

 that antigen is not fixing complement ; test of haemolytic 

 serum and cells and complement with and without antigen 

 to see that haemolysis is actually possible, and tests with 

 known syphilitic serum with and without antigen. In 

 all these controls, except that with known syphilitic serum 

 with antigen, complete haemolysis should occur. 



Noguchi has modified the test by using human red cells 

 and an anti-human haemolytic serum. This simplifies the 

 test in that the patient's red cells may be used in testing 

 his own serum, and in that no antibody for his red cells 

 exists in his own serum (human serum at times contains an 

 antibody to sheep's cells) . Anti-human haemolytic serum is 

 prepared and standardized in a similar way to that 

 described as used in preparing anti-sheep haemolytic serum. 

 Human serum, when used to provide complement, is said 

 to vary more in its content than guinea-pig serum, to absorb 

 10 times as much immune-body, and to be less sensitizing. 



