LANCEFILED GROUPING

This is method for demonstrating precipitation base on reaction of antigen and antibody within the bacteria involved.

Glucose both culture overnight, centrifuge these culture at 3000rmp for 5-10 minute and discard the supernatant also growth culture from blood agar plate can be used.

Add 0.1ml formide to the residue and boil for 1 minute.

Allowed to cool, and add 0.25ml of acid alcohol, boil for 1 minute.

Allowed to cool and add 0.25ml of acid alcohol i.e. HCL hydrochloric acid the purpose of this acid is to clear the fluid by precipitating the protein present in the in the solution, and to precipitate carbohydrate, add acetone.

With a drop of 3 to 4 of saline dissolved the precipitate, phenol red should be added a drop. Used 0.03M NaOH to adjust the pH of the solution to basic contents, centrifuge the solution, this will help in extracting of the supernatant of the solution made up of carbohydrate antigen.

METHOD

On carrying out these test, capillary tube and rack is needed, after the arrangement of capillary tube in the rack that is made up of plastacine, take a loopful of carbohydrate antigen and touch it at the base of the capillary, the antigen will move via capillary attraction immediately, after this take a loopful of antibody and touch the base of the capillary let small amount of the antibody enter the capillary. If it occur the antibody and the antigen are used in the experiment are homogenous there will be rapid precipitation in the capillary within a minute. The test can be carried out with other grouping antiserum.