LYSINS 
167 
serum is determined in the usual manner. The following reagents 
are required to perform the Noguchi test: 
1. Complement— fresh guinea-pig serum in 40 per cent dilution 
(1 part clear fresh serum to 2,5 parts sterile salt solution). 
2. Hemolytic serum— rabbit serum, immunized against human 
erythrocytes and inactivated, is titrated against human erythrocytes 
to determine the hemolytic unit. Two units are used in the test. 
3. Human erythrocytes— red blood cells are obtained from a normal 
individual, washed thoroughly with salt solution, and made up as a 
1 per cent suspension in salt solution. One cubic centimeter of the 
suspension is used in the test. 
4. Antigen— the acetone-insoluble antigen is used. 
5. Patient's serum— obtained fresh, from 2 to 5 cc. of blood. It is 
used unheated. 
6. Known syphilitic serum. 
7. Known normal non-syphilitic serum. 
The test is performed as follows: 
Unknown serum. 
Known positive serum. 
Known negative serum. 
Controls. 
Tube 1. 
Serum, 1 drop 
Complement,' 0.1 cc. 
Erythrocytes, 1.0 cc. 
Tube 3. 
Serum, 1 drop 
Complement, 0.1 cc. 
Erythrocytes, 1.0 cc. 
Tube 5. 
Serum, 1 drop 
Complement, 0.1 cc. 
Erythrocytes, 1.0 cc. 
Tube 7. 
Complement, 0.1 cc. 
Erythrocytes, 1.0 cc. 
Tube 2. 
Serum, 1 drop 
Complement, 0.1 cc. 
Antigen, 2 units 
Erythrocytes, 1.0 cc. 
Tube 4. 
Serum, 1 drop 
Complement, 0.1 cc. 
Antigen, 2 units 
Erythrocytes, 1.0 cc. 
Tube 6. 
Serum, 1 drop 
Complement, 0.1 cc. 
Antigen, 2 units 
Erythrocytes, 1.0 cc. 
Mix and incubate one hour in water-bath at 37° C. Remove and add 2 units hemo- 
lysin to each tube and incubate in water-bath for one hour. Tubes 1, 3 5, 6 and 7 
should show complete hemolysis. Tube 4 should show no hemolysis (positive control). 
If such be the case the reagents are correctly adjusted and a reading of Tube 2 will be 
positive (no hemolysis) or negative (hemolysis) . 
Complement-fixation in Bacterial IniectioTis. — Prejjaration of Antigen 
from /iar/eno.— Experience has clearly shown that bacterial antigens 
should be polyvalent— prepared by mixing several strains of the same 
organism in equal amounts. The antigen may be prepared in one of 
several ways. 
The simplest method is to wash off bacteria from agar slants, at the 
period of maximum growth, with salt solution and shake thoroughly 
to make a uniform suspension. A small amount of phenol (0.5 per 
cent) and 3 per cent glycerin are then added and the whole sterilized 
at 56° to 60° C. for one hour. Relatively more of the proteins of the 
Forty per cent solution of fresh guinea-pig serum in salt solution. 
