404 IMMUNOLOGY 



red cells, and introduced methods for titrating amboceptor and 

 antigen. Wassermann and Bruck employed 1.0 c.c. of a 1 :10 dilu- 

 tion of complement in their test. They also specified that both 

 primary and secondary incubation be carried out at 37° C. The 

 definition for the unit of amboceptor suggested by the Ehrlich 

 school is essentially that employed today. While their final con- 

 centration of red cells was approximately the same as now em- 

 ployed, they did not prepare their suspension from packed cells. 



3. Numerous modifications of the early technique have been 

 used. Noguchi employed an antihuman hemolytic system and a 

 total volume of 1.0 c.c. while the New York City and State Boards 

 of Health have employed a total volume of 0.5 c.c. Others have 

 employed various hemolytic systems and total volumes of 2.5, 3.0, 

 4.0, and 5.0 c.c. The Kolmer technique which is becoming more or 

 less of a standard one in the United States specifies a total volume 

 of 3.0 c.c. 



4. The amount of complement used in the test has varied from 

 0.2 c.c. of undiluted complement employed by Bordet and Gengou, 

 and a titrated amount of a 1 :10 dilution employed by Noguchi to 

 a titrated amount of a 1 :30 employed by Kolmer. 



5. While the time and temperatures employed in the secondary 

 incubation have remained fairly constant since the beginning, the 

 time interval and temperature of the primary incubation have 

 varied. Bordet and Gengou used 15 to 24 hours at 20° C; the 

 Ehrlich school, Noguchi, Craig and others, one hour at 37° C. ; and 

 Kolmer, although not the first to suggest it, has employed 15 to 18 

 hours at 6°-8° C. 



6. The amount of patient's serum employed in the test has 

 varied from 1.2 c.c. suggested by Bordet and Gengou to 0.1 c.c. 

 recommended by Noguchi and also by Kolmer until recently. 

 Kolmer now employs 0.2 c.c. The time of inactivation of serum 

 has varied from 15 minutes to 30 minutes at 56° C. The former 

 time is recommended by Kolmer for the quantitative test and the 

 latter time for the qualitative although he states that 30 minutes 

 may be employed in preparing serum for either test. 



7. Neither Bordet and Gengou nor the Ehrlich school attempted 

 to standardize the glassware employed in the te,st as is done by 



