844 THE COMPLEMENT FIXATION TEST FOR SYPHILIS 



cent cholesterin prepared by a method similar to that of Neymann and Gager. One-tenth of 

 a cubic centimeter of each antigen is used in a dilution that is found to give the most sensitive 

 results in tests of a large number of partially reacting syphilitic sera and that is not even 

 slightly anticomplementary in four times the amount used in the test. 



Four hours at from 3° to 6° C. is allowed for fixation, and with specimens from cases of 

 possible primary syphilis, or those accompanied by histories indicating the necessity for 

 further study, a second test is made with the cholesterinized antigen with fixation in the 

 water bath for one-half hour at 37° C. One-tenth of a cubic centimeter of 5 per cent sheep 

 cells and two units of amboceptor contained in an equal volume are used in the hemolytic 

 system. After the addition of sensitized cells, the tests are incubated at 37° C. for fifteen 

 minutes, or until the serum and antigen controls show complete hemolysis. 



The degree of fixation in each tube is then read by comparison with color standards. 



STANDARDIZATION OF TECHNIQUE 



The desirability of securing some degree of uniformity in the complement fixation 

 test for syphilis has been recognized for years. The solution of the problem has been 

 undertaken through government regulation in both Germany and Great Britain. In 

 the former country the technique for official tests has been prescribed in detail, and 

 strict regulations have been made for the testing of antigens and amboceptor. In 

 Great Britain, the Medical Research Committee,' in 191 8, recommended minimum 

 standards and a choice of four methods which, on the whole, had been found to give 

 satisfactory results. Two years later the British Ministry of Health indorsed the 

 method developed by Griffith and Scott. ^ Representative institutions in these and 

 several other European countries have taken part in the work on serum standardiza- 

 tion of the League of Nations Health Organization. During 1922^ certain precipita- 

 tion reactions were compared with the complement fixation test as performed in the 

 individual laboratories, and during 19234 the different methods for complement fixa- 

 tion also were compared. As a result of these studies there was unanimous agreement 

 that the complement fixation reaction could not be replaced by any of the precipita- 

 tion reactions studied; that the best results were obtained with the complement fixa- 

 tion test when heart extracts were used; and that methods employing several ex- 

 tracts showed no advantage over those employing but one. 



A number of attempts at standardization were also undertaken in this country, 

 some of which were initiated by state and federal authorities. In 191 5 the New York 

 City Department of Health^ made such an attempt, but the undertaking was aban- 

 doned as the members of the committee were unable to agree on technical procedures. 



1 "The Wassermann Test," Rep. of Special Committee upon the Standardization of Patho- 

 logical Methods, "Medical Research Committee Spec. Rep. Ser.," No. 14, p. 270. London, 1918. 



2 Griffith, F., and Scott, W. M.: loc. cit. 



3 "Sero-Diagnosis of Sj^jhilis," Reports on Serological Investigations Presented to the Second Inter- 

 national Conference of the League of Nations Health Organization. Paris, 1922; International Confer- 

 ence on Standardization of Sera and Serological Tests: I. "Sero-Diagnosis of SjTahilis," Lancet, 203, 

 123. 1922. 



"i Investigations on the Serodiagnosis of Syphilis, League of Nations, Health Organization, C. 5 

 M. 5 III. 1924. 



sQttenberg, R.: "On the Reliability of the Wassermann Reaction," Arch. Int. Med., 19, 457. 

 1917. 



