METHODS FOR CONDUCTING THE SYPHILIS REACTION 465 

 TECHNIC OF THE SECOND METHOD 



THE WASSERMANN REACTION WITH MULTIPLE ANTIGENS 



Practically the same technic is used in this as in the first method, 

 except that three different antigens, instead of one, are used with each 

 serum, for the reasons previously stated; for economy the amounts of 

 each reagent are just one-half those originally employed. 



This method is to be strongly recommended, as it is simple, ac- 

 curate, and reliable. Although a little more work is demanded and 

 a larger quantity of the various reagents is required, the results 

 warrant the expenditure of a little more labor, and the second ob- 

 jection is readily overcome by using half the quantities prescribed 

 in the original Wassermann technic, as given in the first method. 



1. I generally use the following three antigens: (1) A cholesterin- 

 ized alcoholic extract of human heart; (2) alcoholic extract of syphil- 

 itic liver; (3) acetone-insoluble lipoids. 



As previously stated, these extracts are used in amounts equal to 

 from two to four times their titrated antigenic unit, providing these 

 doses are at least four times smaller than the anticomplementary 

 units. The amount of each antigen required for the work at hand 

 is calculated, placed in test-tubes, and slowly diluted with the 

 requisite amount of salt solution to secure maximum turbidity of the 

 emulsions. 



2. The complement is diluted 1 : 20 and is used in doses of 1 c.c.; 

 sheep's corpuscles are made up into a 2.5 per cent, suspension, and 

 used in doses of 1 c.c.; antisheep amboceptor is titrated as in the first 

 method, and used in doses equal to 2 units; serums are heated to 

 55 C. for half an hour, and used in doses of 0.1 to 0.2 c.c.; cerebro- 

 spinal fluid is used unheated in amounts of 0.8 c.c. With fresh sera I 

 Ue routinely 0.2 c.c. as the dose; the 0.1 c.c. dose is used in case the 

 amount of serum at hand is insufficient for the 0.2 c.c. dose, or in case 

 the serum is suspected as containing thermostabile anticomplementary 

 substances. 



The complement may be titrated instead of the hemolysin (technic 

 on page 398) and used in a dose equal to 2 units. A large series of 

 comparative tests with both methods and the same sera have yielded 

 almost identical results. 



The Test. For each serum four test-tubes are arranged in a row 



and marked with the patient's name or initials. The first tube is 



marked "C. H.," and receives the cholesterinized heart extract; the 



second tube is marked "S" for the alcoholic extract of syphilitic 



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