Oct. is, 1917 Conglutination Test for the Diagnosis of Glanders 
69 
Table II .—Titration of the conglutination system 
Tube No. 
Comple- 
ment.a 
Inactivated cow serum.* 
Sheep- 
blood cor¬ 
puscles. c 
Result.** 
C. c. 
O. I 
0 . I 
C. c . 
/Cow serum 0.02 1 
[Salt solution 0.78] °' b 
/ Cow serum.. 
[Salt solution 0.8. 
/Cow serum 0.02/ R 
(Salt solution 0.78J 0 * 0 
/Cow serum. 
C. c. 
O. I 
} •« 
. 1 
} -■ 
+ 
e 2 . 
e a 
. 
j 
[Salt solution 0.8 . 
a Fresh normal horse serum. 
* Inactivated cow serum plus salt solution, 
c Five per cent suspension of washed sheep corpuscles. 
+=conglutination; —=no conglutination. 
e Tubes 2, 3, and 4 are controls on the complement, cow serum, and the sheep blood. 
PREPARATION AND TITRATION OF ANTIGEN 
The same antigen that was used in the complement-fixation test was 
used in the conglutination test and is prepared as follows: 
Bacillus mallei is grown for 24 to 48 hours on 5 per cent glycerin agar, 
with the reaction adjusted to H- 2 per cent acid. When the growth has be¬ 
come luxuriant, it is washed off with distilled water carbolized 0.5 per cent 
and then heated at 62° C. for 3 hours. It is then placed in an ice chest 
for four or five days, after which it is centrifuged for 2 hours at a speed 
of 2,100 revolutions per minute. The supernatant fluid, which is quite 
clear, is poured off and represents the antigen. 
A preliminary titration of the antigen is now made with the con¬ 
glutination system. Ten tubes are used, and the. ingredients are added 
in the following order: Antigen, horse complement, inactivated cow 
serum, and sheep blood. The tubes are shaken and allowed to stand at 
room temperature for 2 hours, when the results are read (Table III). 
Table III .—Preliminary titration of the antigen 
Tube No. 
Antigen.** 
Comple¬ 
ment.* 
Inacti¬ 
vated cow 
serum . c 
Sheep- 
blood cor¬ 
puscles.<2 
Result . e 
C . c . 
C.c. 
C.c. 
C.c. 
C . c . 
O.03 
0. I 
0. 8 
0. 1 
+ 
•05 
. I 
.8 
. 1 
+ 
. 08 
. I 
.8 
. 1 
+ 
. I 
. z 
’ .8 
. 1 
+ 
•*5 
. I 
.8 
. 1 
+ 
. 2 
. I 
.8 
. 1 
+ 
•25 
. I 
.8 
. 1 
± 
•3 
. I 
.8 
. 1 
± 
•35 
. I 
.8 
. 1 
— 
• 4 
. I 
. 8 
. 1 
& Dilution of stock antigen 1:10. 
* Fresh normal horse serum. 
c Inactivated cow serum plus salt solution. 
d 0.1 c. c. of a 5 per cent suspension of sheep-blood corpuscles. 
e + ==■ conglutination; impartial conglutination; —=no conglutination. 
