CA1179258A - Integrated method for the determination of anti- streptolysine antibodies and means therefor - Google Patents
Integrated method for the determination of anti- streptolysine antibodies and means thereforInfo
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- CA1179258A CA1179258A CA000389715A CA389715A CA1179258A CA 1179258 A CA1179258 A CA 1179258A CA 000389715 A CA000389715 A CA 000389715A CA 389715 A CA389715 A CA 389715A CA 1179258 A CA1179258 A CA 1179258A
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- streptolysin
- oxidized
- reducing agent
- antibodies
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
- G01N33/56944—Streptococcus
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Abstract
ABSTRACT OF THE DISCLOSURE:
A method is disclosed for determining the anti-streptolysin antibody rating in blood or serum samples, comprising the steps of contacting a solution of known con-centration of oxidized 0-streptolysin with a diluted blood or serum sample, allowing the mixture thus obtained to stand at a temperature of from 15°C to 75°C, adding a solution of a reducing agent, waiting the red blood cell sedimentation, checking if haemolysis occurred and checking the condition of a control blank. If a serum sample is used, it is necessary to add red blood cells thereto prior to the treat-ment with the reducing agent. It is possible to stop the test to the determination of the antibody threshold and to prosecute testing if the preselected threshold has been exceeded. A
diagnostic kit containing the reagents necessary for the tests is also provided.
A method is disclosed for determining the anti-streptolysin antibody rating in blood or serum samples, comprising the steps of contacting a solution of known con-centration of oxidized 0-streptolysin with a diluted blood or serum sample, allowing the mixture thus obtained to stand at a temperature of from 15°C to 75°C, adding a solution of a reducing agent, waiting the red blood cell sedimentation, checking if haemolysis occurred and checking the condition of a control blank. If a serum sample is used, it is necessary to add red blood cells thereto prior to the treat-ment with the reducing agent. It is possible to stop the test to the determination of the antibody threshold and to prosecute testing if the preselected threshold has been exceeded. A
diagnostic kit containing the reagents necessary for the tests is also provided.
Description
S~3 This invention relates to an improved and integrated method for determining the presence and/or the quantity of antistreptolysin antibodies in blood and relates also to the means adapted to such determinations.
As is known, 0-streptolysin is a toxine having a proteinic nature, which is secreted by certain Streptococci of the A Group.
This toxine is lethal for the test animals, it destroys the cell cultures, it lysates the red blood cells and the white blood cells of a number of animal species.
It is likewise known that a few oxidizing agents may succeed to inhibiting the lytic power of such a toxine and, if the oxidation has been gentle, the reaction is reversible:
this means that, under the action of an appropriate reducing agent, the toxine acquires its haemolytic power again.
The determination of the antistreptolysin antibodies has been suggested, long since, as a valid approach for knowing the status and the gravity of a pathological situation connected with the presence of microorganisms which produce 0-streptolysin.
The first procedure to be utilized for this pur-pose has been that provided by TODD (J. Exp. Med., 55, 267-280 (1932): this Author ascertained and showed the presence of antistreptolysin antibodies in the serum of patients affected by infections due to Streptococci. The relative diagnosis test was carried out by neutralizing 0-streptolysin by means of different quantities of the serum being tested, which has previously been deactivated: the excess of non-neutralized 0-streptolysin was evidenced by the haemolysis of the blood red cells added to the system.
RANTZ and RANDALL (Proc. Soc. Exp. Biol. Med., 59, 22-25 (1945)), suggested, in their turn, to utilize, for s~
determining the antistreptolysin rating of the serum, concen-trated solutions of 0-streptolysin which had totally been reduced prior to carrying out the diagnostic test.
The Applicant thereof is the owner of the Canadian Patent N 1.076.025 granted on April 22, 1980 which relates to a particular method for the determination of the antistrepto-lysin rating, a method which provides, inter alia, for carrying out a sequence of steps, the first of which sometimes involves the preparation of a plurality of solutions, having predeter-mined concentrations, of oxidized 0-streptolysin, to which samples of full blood are added, diluted in an appropriate medium and, subsequently, the method provides for the addition of reducing agents which are added to the mixtures as obtained as indicated above, to the end of restoring the haemolytic properties of 0-streptolysin.
- None of the methods indicated heretofore, or any other method as well, has suggested, however, to carry out simultaneously, or in sequential times, according to the desire or the necessity of the operators, a quantitative evaluation of the presence or the absence of the antistreptolysin anti-bodies and/or an accurate determination of the antistreptolysin rating.
The method as described in the above patent even through it is connected with the reliable advantages deriving from the use of 0-streptolysin which has directly been oxidized during progress of the analysis and the use of full blood, is, however, open to criticism, inevitably, whenever it is desired to pass from the accurate determination of the antistreptolysin rating direclty to a purely qualitative estimate of the presence of antibodies only, or of the value of the antibody threshold of a certain sample. As a matter of fact, to resort to a plurality of solutions having different known concentrations of oxidized 0-streptolysin/ affords doubtless many advantages when it is directed to the deter-mination of the 0-antistreptolysin rating, but loses a part of its operational utility when a singl determination of the antibody threshold is desired.
The Applicant hereof has now found, that which is the subject matter of the present invention, a solution which permits to exploit the advantages of the use of the oxidized 0-streptolysin both in the case of the determination of the exact antibody rating and in the case in which it is desired to determine the antibody threshold only.
A first object of the present invention, in fact; is to provide a basic composition which is adapted to achieve both the purposes outlined above, and which is composed of a suitable (i.e. known) quantity of oxidized 0-streptolysin, a reducing agent for restoring the haemolytic properties of 0-streptolysin, a diluent for the sample to be tested, and a control blank. The several components may be held in dif-ferent containers, or, still separate, they can make up a single diagnostic kit.
In both cases, the components may be in a ready-for-use form, or in a concentrated solution, or in solid form, or, also, the oxidized 0-streptolysin can b~ present as a series of solutions of preselected concentrations.
Thus, by way of a nonlimiting example, the composition which is adapted to the determination of the antibodies may be composed of three discrete portions, one of which contains oxidized 0-streptolysin, the other the reducing agent and the third the diluent.
As an alternative, the three portions can be assembled, though separated, in a single kit: oxidized 0-streptolysin, in its turn, may be present in different portions, separated 3~
from each other, in quantities which can be euqal to, or dif-ferent from each other.
Oxidized 0-streptolysin can be present in the solid state, or diluted in an appropriate medium, as a powder, or in any other possible from whatsoever, such as tablets, freeze-dried and otherwise.
A second object of the present invention is to provide a method for the analysis of the antistreptolysin antibodies, such method providing, in its basic formulation, to contact oxidized 0-streptolysin with the sample to be tested, the subsequent treatment with a reducing agent to restore the haemolytic power of the 0-streptolysin, and the comparison with a control blank.
More particularly, the present invention has as its object to provide an integrated method for the analysis of the antistreptolysin antibodies, said method comprising the steps of a preliminary determination of the antibody treshold value and the subsequent accurate measurement of the content of antistreptolysin antibodies: this second step could be dispensed with if the preliminarily obtained value should fall below the threshold limit.
One of the advantages of the method is the possibility of using, indifferently, either full blood or serum, as such and not deactivated.
Detailing now the individual steps, the integrated method for the analysis of the antistreptolysin bodies according to the present invention comprises the steps of preparing a solution of oxidized 0-streptolysin of known concéntrations, preparing one or more solutions of an appropriate reducing agent, diluting the sample with a non-haemolytic medium, contacting the oxidized 0-streptolysin with a preselected quantity of the sample, allowing the mixture thus obtained 5~3 to rest at a temperature comprised between 15C and 75C, adding the reducing agent, waiting for the sedimentation of the red blood cells, controlling the haemolysis or the non-haemolysis condition, and checking the status of the control blank.
The sequence of the several steps outlined above has, of course, its end goal in the determination of the antistrepto-lysin rating, which will be determined as from the last testing step in which no red blood cell haemolysis has been detected.
Such a result can be obtained by starting from solutions of oxidized 0-streptolysin having a constant concentration and adding thereto different quantities of the sample to be tested, or, as an alternative, using a set of solutions of oxidized 0-streptolysin having known concentrations which are gradually increased, and adding to all of them the sample to be tested in the same amount.
In its turn, a single test run carried out by adding a solution of the sample being tested to a solution having a known concentration of 0-streptolysin (oxidized) and subsequently introducing the reducing agent, permits, upon comparison with the control blank, to ascertain the presence, if any, of antistreptolysin antibodies in the sample and to calculate, on the basis of the knowledge of the concentration of 0-streptolysin, the quantity correspondingto such a con-centration. If the concentration is selected properly, this single test run clearly indicates the value of the antibody threshold.
As su~gested hereinbefore, these alternative steps have been made possible by the very nature of the basic composition, which, by providing for the pressure of a number of components, comprises the latter separate from each other, all of them being inserted in the same diagnostic kit, or, if so preferred, each held in its separate container.
3 1'7~
Reducing agents which are adapted to the purpose aremercaptoethanol,dithiothreitol, cysteine, N-acetyl cysteine, sodium metabisulfite, sodium sulfite, just to mention a few.
As outlined above, it is possible to employ, besides full blood, serum: if so, a step of addition of red blood cells must be provided after the addition of the reducing agent to check if haemolysis has taken place, or not.
All the foregoing features and advantages of the invention will become more clearly apparent from the ensuing operative example whereby the invention is to be illustrated without, however, involving any limitation thereto.
EXAMPLE
Assuming to have to carry out the analysis of the antistreptolysin antibodies in a set of 80 blood samples (or serum samples: in such a case it will be necessary to add, at the appropriate time, red blood cells to the individual samples), and assuming, also, that the value of the antibody threshold value has been selected as 200 I.U./ml (International Units per milliliter), a value below which the easy is considered to be negative, while the presence of antistrepto-lysin antibodies above such a value is regarded, conversely, such as to consider the essay positive, there are prepared 80 containers, in each of which oxidized 0-streptolysin is introduced in such an amount as to correspond to the value of the antibody threshold: these containers are the containers for the treshold test.
The 80 blood samples are diluted with an appro-priate diluent and analyzed (according to the procedure outlined above) using the containers for the threshold test.
On completion of the analyses, after having carried out the comparison with the respective control blanks, the samples are divided into two sets, viz.:
1) Samples which did cause haemolysis in their con-tainers: the quantity of antistreptolysin antibodies 0 which is present, is lower than the quantity regarded as the antibody threshold. On these samples the analysis run can be considered as having been completed and the final result is expressed in terms of antistreptolysin rating below 200 I.U.~ml;
As is known, 0-streptolysin is a toxine having a proteinic nature, which is secreted by certain Streptococci of the A Group.
This toxine is lethal for the test animals, it destroys the cell cultures, it lysates the red blood cells and the white blood cells of a number of animal species.
It is likewise known that a few oxidizing agents may succeed to inhibiting the lytic power of such a toxine and, if the oxidation has been gentle, the reaction is reversible:
this means that, under the action of an appropriate reducing agent, the toxine acquires its haemolytic power again.
The determination of the antistreptolysin antibodies has been suggested, long since, as a valid approach for knowing the status and the gravity of a pathological situation connected with the presence of microorganisms which produce 0-streptolysin.
The first procedure to be utilized for this pur-pose has been that provided by TODD (J. Exp. Med., 55, 267-280 (1932): this Author ascertained and showed the presence of antistreptolysin antibodies in the serum of patients affected by infections due to Streptococci. The relative diagnosis test was carried out by neutralizing 0-streptolysin by means of different quantities of the serum being tested, which has previously been deactivated: the excess of non-neutralized 0-streptolysin was evidenced by the haemolysis of the blood red cells added to the system.
RANTZ and RANDALL (Proc. Soc. Exp. Biol. Med., 59, 22-25 (1945)), suggested, in their turn, to utilize, for s~
determining the antistreptolysin rating of the serum, concen-trated solutions of 0-streptolysin which had totally been reduced prior to carrying out the diagnostic test.
The Applicant thereof is the owner of the Canadian Patent N 1.076.025 granted on April 22, 1980 which relates to a particular method for the determination of the antistrepto-lysin rating, a method which provides, inter alia, for carrying out a sequence of steps, the first of which sometimes involves the preparation of a plurality of solutions, having predeter-mined concentrations, of oxidized 0-streptolysin, to which samples of full blood are added, diluted in an appropriate medium and, subsequently, the method provides for the addition of reducing agents which are added to the mixtures as obtained as indicated above, to the end of restoring the haemolytic properties of 0-streptolysin.
- None of the methods indicated heretofore, or any other method as well, has suggested, however, to carry out simultaneously, or in sequential times, according to the desire or the necessity of the operators, a quantitative evaluation of the presence or the absence of the antistreptolysin anti-bodies and/or an accurate determination of the antistreptolysin rating.
The method as described in the above patent even through it is connected with the reliable advantages deriving from the use of 0-streptolysin which has directly been oxidized during progress of the analysis and the use of full blood, is, however, open to criticism, inevitably, whenever it is desired to pass from the accurate determination of the antistreptolysin rating direclty to a purely qualitative estimate of the presence of antibodies only, or of the value of the antibody threshold of a certain sample. As a matter of fact, to resort to a plurality of solutions having different known concentrations of oxidized 0-streptolysin/ affords doubtless many advantages when it is directed to the deter-mination of the 0-antistreptolysin rating, but loses a part of its operational utility when a singl determination of the antibody threshold is desired.
The Applicant hereof has now found, that which is the subject matter of the present invention, a solution which permits to exploit the advantages of the use of the oxidized 0-streptolysin both in the case of the determination of the exact antibody rating and in the case in which it is desired to determine the antibody threshold only.
A first object of the present invention, in fact; is to provide a basic composition which is adapted to achieve both the purposes outlined above, and which is composed of a suitable (i.e. known) quantity of oxidized 0-streptolysin, a reducing agent for restoring the haemolytic properties of 0-streptolysin, a diluent for the sample to be tested, and a control blank. The several components may be held in dif-ferent containers, or, still separate, they can make up a single diagnostic kit.
In both cases, the components may be in a ready-for-use form, or in a concentrated solution, or in solid form, or, also, the oxidized 0-streptolysin can b~ present as a series of solutions of preselected concentrations.
Thus, by way of a nonlimiting example, the composition which is adapted to the determination of the antibodies may be composed of three discrete portions, one of which contains oxidized 0-streptolysin, the other the reducing agent and the third the diluent.
As an alternative, the three portions can be assembled, though separated, in a single kit: oxidized 0-streptolysin, in its turn, may be present in different portions, separated 3~
from each other, in quantities which can be euqal to, or dif-ferent from each other.
Oxidized 0-streptolysin can be present in the solid state, or diluted in an appropriate medium, as a powder, or in any other possible from whatsoever, such as tablets, freeze-dried and otherwise.
A second object of the present invention is to provide a method for the analysis of the antistreptolysin antibodies, such method providing, in its basic formulation, to contact oxidized 0-streptolysin with the sample to be tested, the subsequent treatment with a reducing agent to restore the haemolytic power of the 0-streptolysin, and the comparison with a control blank.
More particularly, the present invention has as its object to provide an integrated method for the analysis of the antistreptolysin antibodies, said method comprising the steps of a preliminary determination of the antibody treshold value and the subsequent accurate measurement of the content of antistreptolysin antibodies: this second step could be dispensed with if the preliminarily obtained value should fall below the threshold limit.
One of the advantages of the method is the possibility of using, indifferently, either full blood or serum, as such and not deactivated.
Detailing now the individual steps, the integrated method for the analysis of the antistreptolysin bodies according to the present invention comprises the steps of preparing a solution of oxidized 0-streptolysin of known concéntrations, preparing one or more solutions of an appropriate reducing agent, diluting the sample with a non-haemolytic medium, contacting the oxidized 0-streptolysin with a preselected quantity of the sample, allowing the mixture thus obtained 5~3 to rest at a temperature comprised between 15C and 75C, adding the reducing agent, waiting for the sedimentation of the red blood cells, controlling the haemolysis or the non-haemolysis condition, and checking the status of the control blank.
The sequence of the several steps outlined above has, of course, its end goal in the determination of the antistrepto-lysin rating, which will be determined as from the last testing step in which no red blood cell haemolysis has been detected.
Such a result can be obtained by starting from solutions of oxidized 0-streptolysin having a constant concentration and adding thereto different quantities of the sample to be tested, or, as an alternative, using a set of solutions of oxidized 0-streptolysin having known concentrations which are gradually increased, and adding to all of them the sample to be tested in the same amount.
In its turn, a single test run carried out by adding a solution of the sample being tested to a solution having a known concentration of 0-streptolysin (oxidized) and subsequently introducing the reducing agent, permits, upon comparison with the control blank, to ascertain the presence, if any, of antistreptolysin antibodies in the sample and to calculate, on the basis of the knowledge of the concentration of 0-streptolysin, the quantity correspondingto such a con-centration. If the concentration is selected properly, this single test run clearly indicates the value of the antibody threshold.
As su~gested hereinbefore, these alternative steps have been made possible by the very nature of the basic composition, which, by providing for the pressure of a number of components, comprises the latter separate from each other, all of them being inserted in the same diagnostic kit, or, if so preferred, each held in its separate container.
3 1'7~
Reducing agents which are adapted to the purpose aremercaptoethanol,dithiothreitol, cysteine, N-acetyl cysteine, sodium metabisulfite, sodium sulfite, just to mention a few.
As outlined above, it is possible to employ, besides full blood, serum: if so, a step of addition of red blood cells must be provided after the addition of the reducing agent to check if haemolysis has taken place, or not.
All the foregoing features and advantages of the invention will become more clearly apparent from the ensuing operative example whereby the invention is to be illustrated without, however, involving any limitation thereto.
EXAMPLE
Assuming to have to carry out the analysis of the antistreptolysin antibodies in a set of 80 blood samples (or serum samples: in such a case it will be necessary to add, at the appropriate time, red blood cells to the individual samples), and assuming, also, that the value of the antibody threshold value has been selected as 200 I.U./ml (International Units per milliliter), a value below which the easy is considered to be negative, while the presence of antistrepto-lysin antibodies above such a value is regarded, conversely, such as to consider the essay positive, there are prepared 80 containers, in each of which oxidized 0-streptolysin is introduced in such an amount as to correspond to the value of the antibody threshold: these containers are the containers for the treshold test.
The 80 blood samples are diluted with an appro-priate diluent and analyzed (according to the procedure outlined above) using the containers for the threshold test.
On completion of the analyses, after having carried out the comparison with the respective control blanks, the samples are divided into two sets, viz.:
1) Samples which did cause haemolysis in their con-tainers: the quantity of antistreptolysin antibodies 0 which is present, is lower than the quantity regarded as the antibody threshold. On these samples the analysis run can be considered as having been completed and the final result is expressed in terms of antistreptolysin rating below 200 I.U.~ml;
2) Samples which did not cause haemolysis in their containers: the quantity of antistreptolysin antibodies 0 which is present is equal to, or higher than the quantity regarded as being the antibody threshold. On these samples the analysis run cannot be regarded as having been completed since it is deemed necessary to know the exact antistreptolysin rating.
Still considering the example of the analysis of 80 samples, let it be assumed that also the samples of the second group are 10 in number. There are thus prepared 10 sets of containers which contain gradually increasing and predetermined quantities of oxidized 0-streptolysin (the number of the containers of each set and the quantities of oxidized 0-streptolysin are selected consistently with the range of antistreptolysin rating one desires to assay).
The samples in question are then analyzed (accord-ing the procedure as described hereinbefore), each with a set of containers.
The final result is defined by the quantity of 0-streptolysin which is present in the container in which no red blood cell haemolysis has been detected and which immediately preceeds the first of the containers in which red blood cell haemolysis has been detected.
Still considering the example of the analysis of 80 samples, let it be assumed that also the samples of the second group are 10 in number. There are thus prepared 10 sets of containers which contain gradually increasing and predetermined quantities of oxidized 0-streptolysin (the number of the containers of each set and the quantities of oxidized 0-streptolysin are selected consistently with the range of antistreptolysin rating one desires to assay).
The samples in question are then analyzed (accord-ing the procedure as described hereinbefore), each with a set of containers.
The final result is defined by the quantity of 0-streptolysin which is present in the container in which no red blood cell haemolysis has been detected and which immediately preceeds the first of the containers in which red blood cell haemolysis has been detected.
Claims (13)
1. A composition suitable for the analysis of the antistreptolysin antibodies, composed of a suitable amount of oxidized O-streptolysin, a reducing agent for restoring the haemolytic power of O-streptolysin, a diluent for the sample to be tested and a control blank.
2. Composition according to Claim 1, wherein the oxidized O-streptolysin is present in a subdivided set of samples in preselected concentrations.
3. A diagnostic kit for the analysis of the antistreptolysin antibodies, consisting, in separate containers, increasing quantities of oxidized O-strepto-lysin in known amounts, a reducing agent for restoring the haemolytic properties of O-streptolysin and a diluent for the sample to be tested.
4. Diagnostic kit according to Claim 3, wherein the oxidized O-streptolysin in known amounts is present in a set of separate containers each containing a pre-selected concentration of oxidized O-streptolysin said kit including a control blank.
5. An integrated method for the analysis of the antistreptolysin antibodies comprising the step of preliminarily determining the threshold value of the antibody content of a sample and subsequently, if desired, accurately measuring the antistreptolysin antibody rating, said preliminary step being carried out by con-tacting a known quantity of oxidized O-streptolysin with the sample concerned, said sample, if desired, being diluted with a non-haemolytic medium, subsequently treating what has been obtained with a reducing agent for restoring the haemolytic properties of O-streptolysin, waiting the red blood cell sedimentation, checking the possible haemolysis of the red cells and checking the status of a control blank.
6. Integrated method for the analysis of the anti-streptolysin antibodies according to claim 5, comprising the steps of preparing one or more solutions of known concen-trations of oxidized O-streptolysin, preparing a solution of an appropriate reducing agent, diluting the sample to be tested with a non-haemolytic medium, contacting the oxidized O-streptolysin with a preselected amount of the sample to be tested, allowing the mixture thus obtained to stand at a temperature comprised between 15°C and 75°C, adding a reducing agent, waiting the red blood cell sedimentation, checking haemolysis and checking the conditions of the control blank.
7. Integrated method for the analysis of the anti-streptolysin antibodies according to either one of claims 5 and 6, wherein the sample to be tested is full blood or serum as such.
8. Integrated method for the analysis of the anti-streptolysin antibodies according to either one of claims 5 and 6, wherein the sample to be tested is serum as such and the restoration of the haemolytic properties of O-streptolysin is preceded by an appropriate addition of red blood cells.
9. A composition according to claim l or claim 2, wherein oxidized O-streptolysin is present in the solid state, or in solution at known concentration, or in the freeze-dried state.
10. Composition according to claim l or claim 2, wherein the reducing agent is a member selected from the group consisting of mercaptoethanol, dithiothreitol, cysteine, N-acetylcysteine, sodium metabisulfite and sodium sulfite.
11. A diagnostic kit according to claim 3 or claim 4, wherein oxidized O-streptolysin is present in the solid state, in solutions of known concentrations or also in the freeze-dried state.
12. A diagnostic kit according to claim 3 or claim 4, wherein the reducing agent is a member selected from the group consisting of mercaptoethanol, dithiothreitol, cysteine, N-acetylcysteine, sodium metabisulfite and sodium sulfite.
13. An integrated method for the combined detection and quantitive measurement of antistreptolysin antibodies, said method comprising, (a) diluting a plurality of samples of whole blood with a non-haemolytic medium;
(b) contacting said samples with an amount of oxidized O-streptolysin which corresponds to the thres-hold of the antibody;
(c) allowing the mixtures thus obtained to rest at a temperature between 15°C and 75°C;
(d) adding a reducing agent for restoring the haemolytic properties of O-streptolysin to the mixtures of step (c);
(e) selecting the samples that did not exhibit haemolysis and identifying those samples as containing antistreptolysin antibodies;
(f) adding the samples of step (e) that did not exhibit haemolysis to a plurality of containers containing increasing and known quantities of oxidized O-streptolysin;
(g) adding a reducing agent for restoring the haemolytic properties of O-streptolysin to each of the containers of step ( f);
(h) determining the quantity of antistrep-tolysin antibodies that are present by selection of the container containing the maximum quantity of O-strepto-lysin that does not cause haemolysis.
(b) contacting said samples with an amount of oxidized O-streptolysin which corresponds to the thres-hold of the antibody;
(c) allowing the mixtures thus obtained to rest at a temperature between 15°C and 75°C;
(d) adding a reducing agent for restoring the haemolytic properties of O-streptolysin to the mixtures of step (c);
(e) selecting the samples that did not exhibit haemolysis and identifying those samples as containing antistreptolysin antibodies;
(f) adding the samples of step (e) that did not exhibit haemolysis to a plurality of containers containing increasing and known quantities of oxidized O-streptolysin;
(g) adding a reducing agent for restoring the haemolytic properties of O-streptolysin to each of the containers of step ( f);
(h) determining the quantity of antistrep-tolysin antibodies that are present by selection of the container containing the maximum quantity of O-strepto-lysin that does not cause haemolysis.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| IT21549/81A IT1218273B (en) | 1981-05-07 | 1981-05-07 | INTEGRATED METHOD FOR ANALYSIS OF ANTI-STREPTOLYSINIC ANTIBODIES AND MEDIA SUITABLE FOR THE PURPOSE |
| IT21549A/81 | 1981-05-07 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1179258A true CA1179258A (en) | 1984-12-11 |
Family
ID=11183447
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000389715A Expired CA1179258A (en) | 1981-05-07 | 1981-11-09 | Integrated method for the determination of anti- streptolysine antibodies and means therefor |
Country Status (18)
| Country | Link |
|---|---|
| AR (1) | AR229167A1 (en) |
| AU (1) | AU551120B2 (en) |
| BE (1) | BE891075A (en) |
| BR (1) | BR8107026A (en) |
| CA (1) | CA1179258A (en) |
| CH (1) | CH655801A5 (en) |
| DE (1) | DE3144042A1 (en) |
| DK (1) | DK470581A (en) |
| ES (1) | ES8302310A1 (en) |
| FI (1) | FI813512L (en) |
| FR (1) | FR2505502B1 (en) |
| GB (1) | GB2098322B (en) |
| GR (1) | GR76695B (en) |
| IT (1) | IT1218273B (en) |
| NO (1) | NO813772L (en) |
| NZ (1) | NZ198700A (en) |
| PT (1) | PT73958B (en) |
| SE (1) | SE8106638L (en) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TR19262A (en) * | 1975-12-22 | 1978-09-19 | Sclavo Inst Sieroterapeut | TITRATION PROCEDURE MADE BY THE DEVICE AND THE ELEMENTS OF THIS DEVICE FOR TITRATION OF ANTISTREPTOLINE ANTIBODIES |
| IT1132167B (en) * | 1980-07-03 | 1986-06-25 | Diesse Diagnostica Senese Srl | HEMOLYTIC METHOD FOR THE CONTINUOUS DETERMINATION OF ANTI-STREPTOLYSINE ANTIBODIES OR, IN BLOOD OR SERUM SAMPLES WITH THE USE OF STREPTOLYSINE OR OXIDIZED |
-
1981
- 1981-05-07 IT IT21549/81A patent/IT1218273B/en active
- 1981-10-20 NZ NZ198700A patent/NZ198700A/en unknown
- 1981-10-26 GR GR66356A patent/GR76695B/el unknown
- 1981-10-26 DK DK470581A patent/DK470581A/en not_active Application Discontinuation
- 1981-10-26 BR BR8107026A patent/BR8107026A/en unknown
- 1981-11-02 AU AU77045/81A patent/AU551120B2/en not_active Ceased
- 1981-11-05 DE DE19813144042 patent/DE3144042A1/en not_active Withdrawn
- 1981-11-09 GB GB8133724A patent/GB2098322B/en not_active Expired
- 1981-11-09 SE SE8106638A patent/SE8106638L/en unknown
- 1981-11-09 FI FI813512A patent/FI813512L/en not_active Application Discontinuation
- 1981-11-09 NO NO813772A patent/NO813772L/en unknown
- 1981-11-09 AR AR287389A patent/AR229167A1/en active
- 1981-11-09 CA CA000389715A patent/CA1179258A/en not_active Expired
- 1981-11-09 FR FR8120974A patent/FR2505502B1/en not_active Expired
- 1981-11-10 PT PT73958A patent/PT73958B/en unknown
- 1981-11-10 CH CH7224/81A patent/CH655801A5/en not_active IP Right Cessation
- 1981-11-10 BE BE0/206510A patent/BE891075A/en not_active IP Right Cessation
- 1981-11-10 ES ES507495A patent/ES8302310A1/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| DE3144042A1 (en) | 1982-11-25 |
| AR229167A1 (en) | 1983-06-30 |
| NZ198700A (en) | 1985-12-13 |
| FR2505502A1 (en) | 1982-11-12 |
| GR76695B (en) | 1984-08-24 |
| BE891075A (en) | 1982-05-10 |
| ES507495A0 (en) | 1982-12-16 |
| CH655801A5 (en) | 1986-05-15 |
| PT73958B (en) | 1983-11-23 |
| DK470581A (en) | 1982-11-08 |
| ES8302310A1 (en) | 1982-12-16 |
| FR2505502B1 (en) | 1985-08-23 |
| IT1218273B (en) | 1990-04-12 |
| GB2098322A (en) | 1982-11-17 |
| BR8107026A (en) | 1983-04-12 |
| IT8121549A0 (en) | 1981-05-07 |
| NO813772L (en) | 1982-11-08 |
| SE8106638L (en) | 1982-11-08 |
| FI813512L (en) | 1982-11-08 |
| PT73958A (en) | 1981-12-01 |
| AU7704581A (en) | 1982-11-11 |
| AU551120B2 (en) | 1986-04-17 |
| GB2098322B (en) | 1985-02-06 |
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