Rivanol Brucella Antigen and Rivanol Solution:
 


COMPOSITION :
The antigen is prepared from a concentrated cell suspension of CO2 independent smooth strain of Br.abortus (strain 99). The cells are stained with brilliant green and crystal violet stain and suspended in citrated buffer diluent with final pH 5.8 – 6.2 .
Rivanol solution is a 1% (w/v) solution of Rivanol powerd (2- ethoxy – 6.9 – diamine – acridine lactate).


 
 
 
 


INDICATION :
It is used for serological diagnosis of brucellosis in cattle and sheep Rivanol plate test Brucella antigen. It is a supplementary test. The Rivanol solution selectively precipitates IgM in the serum, this then removed by centrifugation and the supernatant is tested for identification of agglutinating IgG. This test has advantage of high specificity.


 
 
 
 


METHOD OF USE :
The Rivanol plate test is run at 4 dilutions correspond to the 1:25, 1:50, 1:100 and 1:200 dilutions of the USDA standard tube test.
1- Allow the serum and reagents to warm to room temperature for at least 1 hour.
2- With a 1 ml pipette, measure 0.4 ml of Rivanol solution into the required number of suitably sized (say 1 or 14×100 mm) tubes.
3- Using a fresh 1.0 ml pipette, for each sample, measure 0.4 ml of serum under test into each tube and mix immediately by shaking the tube.
4- Leave the tubes at room temperature for at least 5 minutes but not more than 1 hour, then centrifuge them at approximately 1000g. for 5 minutes or at a lesser g. force for long enough to pack the precipitate and leave a clear yellow supernatant.
5- Using a 0.2 ml pipette, measure 0.08, 0.04, 0.02, 0.01 ml quantities of supernatant fluid on to separate squares of the glass plate of the testing box.
6- Add one drop(0.03 ml) of Rivanol antigen to each quantity of supernatant fluid, mix with a stirrer, starting with the smallest quantity (0.01 ml). Each quantity should be spread to approximately an area 2 cm. in diameter for 0.01 serum drop (1:200 dil) increasing the diameter of the spread progressively up to about 3 cm. for the 0.08 ml serum drop (1:25
dilution).
7- Rinse and dry the stirrer between samples.
8- Tilt the plate in a circular motion for 4 rotations .
9- Replace the plate in the box and close the lid to prevent evaporation.
10- After 6 minutes again rotate the plate as before for 4 rotations.
11- After another 6 minutes again rotate the plate for 4 rotation and read the reaction against the illuminated background of the box .
12- The test is read and the results recorded.

N.B. For convenience the mixtures of supernatant and antigen are taken as 1:25, 1:50, 1:100, 1:200 as the USDA standard tube test .


 
 
 
 


INTERPERATION :
In non – vaccinated cattle, complete agglutination at 1:25 should be considered positive. In clashed vaccination complete agglutination at 1:25 may be considered as suspicious, and complete agglutination at 1:50 as positive.


  
 
 
 


PRECAUTIONS :
* Shake the antigen very well before use.
* To ensure validity of the antigen use control positive and negative sera.

 

  
 
 


STORAGE :
Avoid freezing – keep the antigen at 4 to 8oC.

 
 
 
 


EXPIARY DATE :
One year from the date of manufacture.

 

  
 
 
 


PACKAGE :
Glass bottle of 20 ml antigen to apply 600 tests.
Glass bottle of 60 ml 1% Rivanol solution.