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TECHNIQUE OF THE COMPLEMENT FIXATION METHOD 



-f- complement (five hours) + hemolysin + blood. Hemolysis must occur, as the 

 normal serum does not contain enough amboceptors to unite with the bacterial suspen- 

 sion and consequently complement remains unbound. 



2. Inactivated immune serum + complement (five hours) + hemolysin + blood. 

 Hemolysis results. 



3. Inactivated normal serum + complement (five hours) + hemolysin + blood. 

 Hemolysis. 



4. Antigen + inactivated immune serum + hemolysin + blood. No hemolysis, 

 as complement is absent. 



5. Antigen + inactivated normal serum (five hours) + hemolysin -f- blood. No 

 hemolysis, as complement is absent. 



The following is the chart of the first complement fixation test as originally per- 

 formed by Bordet and Gengou in 1901 in which pest antibodies were demonstrated in 

 the serum of an immunized horse. 



Employing this method, Bordet and Gengou found positive results with the 

 following combinations: 



1 . Pest bacilli + pest horse's serum + guinea-pig complement -f- guinea-pig hem- 

 olysin -t- rabbit's blood. 



2. Anthrax vaccine + guinea-pig immune serum + guinea-pig complement -f- 

 guinea-pig hemolysin -f rabbit's blood. 



3. Typhoid bacilli + guinea-pig immune serum + guinea-pig complement + 

 guinea-pig hemolysin + rabbit's blood. 



4. Coli bacilli + guinea-pig immune serum + guinea-pig complement + guinea- 

 pig hemolysin + rabbit's blood. 



5. Typhoid bacilli -f- human convalescent serum + human complement + guinea- 

 pig hemolysin + rabbit's blood. 



