The complement fixation test in syphilis, commonly known as the Wassermann test.

  • Great Britain. Ministry of Health.
Date:
1920
Catalogue details

Licence: Public Domain Mark

Credit: The complement fixation test in syphilis, commonly known as the Wassermann test. Source: Wellcome Collection.

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    ) Ingredients.—Normal saline prepared by dissolving 0°85 per cent. chemically pure sodium chloride in fresh glass-distilled water followed by filtration and sterilisation at 120° C. in the autoclave. Human serum diluted 1 in 5 with normal saline and then heated for half an hour at 56° C. Complement obtained by bleeding male guinea-pigs and used the same day, diluted 1 in 25 with normal saline. Antigen prepared by adding normal saline quickly to alcoholic solution of acetone-insoluble lipoids to which alcoholic solution of cholesterin has been added in the requisite amount ; the resulting suspension of lipoids should be clear or faintly opalescent, and should contain 1 part of the mixture of alcoholic solutions to 640 parts of normal saline. Blood cells obtained from sheep’s blood, collected in citrate solution, washed free from serum, formolised and made up with normal saline so that 6 per cent. ‘of the sediment is present in the suspension. Hemolytic immune serum obtained from horses injected with sheep’s blood cells. Sensitised blood cell suspension prepared by mixing equal quantities of the 6 per cent. blood cell suspension and normal saline containing sufficient of the hemolytic immune serum to give maximum sensitisation. Order of Work.—Preliminary Steps :— (1) Human sera to be pipetted from clot, centrifuged if necessary, then 0°25 ¢.c. to be mixed with 1 c.c. of normal saline and heated for half an hour at 56° C. in the water-bath. (2) Guinea-pig to be bled, serum pipetted off, centrifuged, and diluted 1 in 25 with normal saline. (3) Sensitised blood cell suspension prepared and kept at room T” in sufficient amount for the preliminary titration, e.g., 0°25 c.c. sediments+ 3°9 c.c. normai saline + 4°*le.c. of the dilution of hemolytic immune serum. . (4) Preliminary titration of complement (to ensure that it is of normal lytic activity). (5) Preparation of antigen suspension maintained at. 0° C. in ice-bath. Setting up of Test and Controls. Fixation stage :— Arrangement of tubes. (a) Three tubes for each serum to be set one behind the other, and two sets of four tubes for the complement controls. Addition of diluted human serum. (b) Add 0°25 c.c. normal saline to each tube of the third row, then 0°25 cc. of the appropriate diluted human serum to each of the three tubes arranged for its test. Remove 0°25 c.c. of the mixture from