Method for measuring antistreptolysin concentration in blood

Abstract

A method for measuring the antistreptolysin concentration in a human blood sample by adding a solution of oxidized O-streptolysin to the blood sample (diluted), incubating, reacting with a reducing solution, incubating and relating the resulting hemolysis to the antistreptolysin concentration.

What I claim is: 
     
       1. A method for measuring the antistreptolysin concentration in a sample of human blood, comprising the steps of: (a) preparing a plurality of solutions of different concentrations of oxidized O-streptolysin;   (b) preparing a solution of a reducing substance selected from the group consisting of thiols, sodium metabisulfite and sodium sulfite;   (c) diluting a blood sample with a nonhemolytic solution;   (d) introducing an equal volume of diluted blood in each of the solutions prepared in step (a);   (e) incubating the solutions prepared in step (d) at a temperature in the range between room temperature (18° C.-21° C.) and 50° C. for about 15 minutes;   (f) adding to the incubating solutions prepared in step (e) a measured volume of the solution prepared in step (b);   (g) incubating the solution prepared in step (f) at a temperature in the range between room temperature and 50° C. for sufficient time to cause the sedimentation of the red blood cells to occur, and   (h) noting the sample containers in which no hemolysis has taken place. .Iadd.   
     
     
       2.  In a method for determining whether a detectable quantity of antistreptolysin is present in human blood, the steps which comprise: (A) preparing a solution of a preselected concentration of oxidized O-streptolysin,   (B) preparing a solution of a reducing substance selected from the group consisting of thiols, sodium metabisulfite and sodium sulfite,   (C) preparing a blood sample for determination,   (D) introducing a measured quantity of said blood sample into the solution prepared in step (A),   (E) incubating the solution prepared in step (D) at a temperature and for a sufficient time to allow for the neutralization of the oxidized O-streptolysin should a detectable quantity of antistreptolysin be present in said blood,   (F) adding to the incubating solution prepared in step (E) a measured volume of the solution prepared in step (B),   (G) incubating the solution prepared in step (F) at a temperature and for a sufficient time to cause the sedimentation of the red blood cells to occur, and   (H) noting whether or not hemolysis has taken place therein. .Iaddend. .Iadd.3. The method of claim 2 wherein the incubation of the solution recited in step (E) is carried out at a temperature in the range between room temperature (18 deg. C.-21 deg. C.) and about 50 deg. C. for about 15   
     
     
        minutes. .Iaddend. .Iadd.4.  The method of claim 2, wherein the incubation of solution recited in step (G) is carried out at a temperature in the 
     
     
        range between room temperature and 50 deg. C. .Iaddend. .Iadd.5.  The method described in claim 2 wherein the incubation of the solution recited in step (e) is carried out at a temperature in the range between room temperature (18° C. -21° C.) and about 50° C. for about 15 minutes, and   wherein the incubating of the solution recited in step (f) is carried out at a temperature in the range between room temperature and 50° C. .Iaddend.