BACTERIAL COMPLEMENT FIXATION TECHNIQUE 413 



occur if tlie antigen is not anticomplementary. It is suggested that 

 the student follow Kolmer's suggestions. 



Quantitative and Qualitative Tests. — The technique of the 

 quantitative test is similar to that of the simplified test except 

 that in the former the serum is used in amounts of 0.2, 0.1, 0.05, 

 0.025, and 0.005 c.c. with 0.2 c.e. in the control tube. In the 

 qualitative test Kolmer uses 0.2 and 0.1 c.c. and 0.2 c.c. in the 

 control. Positive, negative and other controls should be included in 

 both tests. 



Reading the Results. — Kolmer states that the readings should 

 be made immediately after the secondary incubation of one hour 

 or they may be made 10 minutes after complete hemolysis occurs 

 in the serum, antigen and hemolytic system controls. 



Reporting- Results. — Kolmer states that results may be reported 

 as ''positive," ''doubtful" or "negative." He considers fixation 

 of one plus or more as ' ' positive ' ' ; the + reactions are called 

 "doubtful" and when complete hemolysis occurs the report is 

 "negative." Not infrequently the physician desires to know the 

 degree of fixation observed. In such an event the report is given 

 as four, three, two or one plus positive or + as doubtful and 

 complete hemolysis as negative fixation. 



Correlation With Clinical Findings. — In the case of gonococcal 

 complement fixation, Price obtained 85 per cent positive results 

 in known cases of infection. Park, AVilliams and Krumwiede 

 re])ort 75 per cent to 95 per cent positive results for complement 

 fixation in clinically active pulmonary tuberculosis. In glandular 

 tuberculosis and tuberculosis of the l)ones and joints, only 58 

 per cent and 22 per cent, respectively, of the sera gave positive 

 results using complement fixation. 



Discussion. — It should be remembered that the complement 

 fixation test is just one of several techniques employed to detect 

 antibody in blood serum and it is perhaps one of the most sensi- 

 tive tests employed for that purpose. The reason for lack of cor- 

 relation with clinical findings may be due to : 



1. Defective reagents such as antigen, complement, hemolysin 

 and saline. 



2. Defective technique in preparing glassware or reagents, or 

 errors in inactivation, incubation, or in setting up the test, 

 «tc. 



3. The serum being tested. 



