BACTERIAL COMPLEMENT FIXATION TECHNIQUE 405 



Koliner. There are, however, good reasons for adopting standard 

 equipment and standardized procedures. The pipettes he recom- 

 mends enable the technician to combine accuracy with reasonable 

 speed. The test tubes are of dimensions to insure a satisfactory 

 height to the column of content.s and permit adequate mixing of 

 reagents by shaking the rack. There are equally good reasons 

 for most of his other requirements. 



8. In regard to the diluent used (0.85 per cent saline) in com- 

 plement fixation work, it has remained essentially the same since 

 its introduction by Bordet and Gengou with the exception that 

 Kolmer adds 0.10 gm. of magnesium sulphate per liter. Since 

 Kolmer has increased the sensitivity of his modification of the 

 Bordet- Wassermann test by reducing the concentration of com- 

 plement and by increasing the lipoid content of his antigens and 

 by interpreting a one-plus reaction as positive, there has been a 

 growing insistence upon the use of purer saline solutions. 



9. As regards bacterial antigens to be employed in complement 

 fixation considerable progress has been made but there is room for 

 improvement. Bordet and Gengou used untreated bacterial sus- 

 l)ensions Avhile Wassermann and Bruck used bacterial extracts. 

 Since then the New York City Board of Health has employed 

 polyvalent antigens defatted with alcohol and ether; Wadsworth 

 (1929) describes a "Dialyzed Distilled Water-Extract" method 

 for the preparation of an antigen from tubercle bacilli ; Price 

 (1932) dissolves a suspension of the gonococcus in NaOH and 

 later adds sufficient HCl to precipitate out an antigenic substance 

 and Kolmer and Boerner (1941) describe other methods of bac- 

 terial antigen preparation. Torrey (1940) recommends Price's 

 antigen for gonococcal complement fixation. 



Theoretically every antigen should l)e tested to see if it is 

 hemolytic, anticomplementary or antigenic, but Kolmer (1941) says 

 that when the bacterial antigens are prepared according to his 

 method they are rarely hemolytic and therefore the hemolytic 

 titration may be safely omitted. For the sake of completeness we 

 will include protocols of the hemolytic, anticomplementary and 

 antigenic titrations as recommended by Kolmer. They are as 

 follows ; 



