840 THE COMPLEMENT FIXATION TEST FOR SYPHILIS 



METHOD USED BY BROWNING, AT THE BLAND-SUTTON INSTITUTE, MIDDLESEX HOSPITAL; 



ONE OF THE FOUR METHODS RECOMMENDED IN 1918 BY THE MEDICAL RESEARCH 



COMMITTEE OF GREAT BRITAIN' 



The patient's serum is inactivated for one-half hour at 56° C, and 0.025 cc. is used in the 

 test. 



Guinea pig serum collected eighteen hours previously is used as complement. It is 

 titrated in a 1:4 dilution with incubation for one hour at 37° C. Two, four, six, and eight 

 units are used with constant amounts of patient's serum and antigen. 



The antigens are alcoholic "lecithin" solutions of ox liver used with and without the addi- 

 tion of cholesterin. The optimum amount of cholesterin is determined by testing different 

 concentrations in comparison with an antigen that has proved to be satisfactory and is in 

 routine use. Three-tenths of a cubic centimeter of each antigen in a 1:8 dilution are used in 

 the tests. Precautions are taken in making the dilution to insure minimum turbidity. 



Anti-ox amboceptor is titrated with a 3 per cent suspension of cells, and five units are 

 used in the hemolytic system. 



One and one-half hours at 37° C. are allowed for fixation, and after the addition of ambo- 

 ceptor and cells, there is a secondary incubation at 37° C. for one hour. 



The results are evaluated by comparison with the reactions obtained with specimens 

 that give varying degrees of fixation and are tested for purposes of control. 



METHOD OF GRIFFITH AND SCOTT^ 



All the components of the test are diluted so that 0.25 cc. of each are used. The patient's 

 serum is diluted i : 5 with physiological salt solution and then inactivated for one-half hour 

 at 56° C. Both 1 : 5 and i : 10 dilutions are used in the test. 



The complement consists of 0.25 cc. of a 1:25 dilution of the serum from male guinea 

 pigs that has been collected on the day the tests are made. It is tested both with and without 

 the addition of antigen to control its lytic activity. 



As antigen, an acetone-insoluble fraction of crude organ extract as suggested by Noguchi 

 is recommended. To this is added an equal volume of alcohol, containing i per cent choles- 

 terin. When diluting, six hundred and forty parts of physiological salt solution aie added 

 rapidly to one part of the mixture of alcoholic solutions, so that the resulting suspension is 

 clear or faintly opalescent. Patient's serum and complement are mixed and allowed to stand 

 for one hour in the icebox after which the cool antigen suspension is added. They are then 

 replaced in the icebox fcr from sixteen to twenty-four hours. For the hemolytic system, 6 

 per cent suspension of sheep cells and an equal volume of antisheep horse serum in a dilution 

 sufticient to furnish maximum sensitization is used. 



Reactions are recorded, depending upon the degree of lysis, as follows: o, tr, ?tr, +, 

 + + ,+ + + ,+ + + + , ?C, and C. 



METHOD USED BY MADSEN, STATE SERUM INSTITUTE, COPENHAGEN' 



The technique indicated by Thomsen and Boas with an additional antigen prepared from 

 syphilitic liver is used. The serum is inactivated for one-half hour at 56° C, and amounts of 



' "The Wassermann Test," Rep. of Special Committee upon the Standardization of Pathological 

 Methods, "Medical Research Committee Spec. Rep. Ser.," No. 14, p. 27. London, 1918. 



^ Griffith, F., and Scott, W. M.: //. Technique of the Wassermann Reaction, Rep. on Pub. Health 

 and Med. Subjects, No. i, p. 7. London: Ministry of Health, 1920. 



3 Investigations on the Serodiagnosis of Syphilis, League of Nations, Health Organization, C. 5 M. 5 

 III. 1924. 



