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US5091304A
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Whole blood activated partial thromboplastin time test and associated apparatus
Assignee: International Technidyne Corp
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Filing: 1989-08-21
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Granted: 1992-02-25
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Expires: 2009-08-21
Abstract
Abstract An activated partial thromboplastin time (APTT) test is described which does not require blood which has been anticoagulated with citrate. The test enables the citrate anticoagulant step to be combined with the contact activation step and hence enables one to employ fresh non-anticoagulated blood specimens to directly perform the APTT Test.
Inventors
- 👤 Frank M. La Duca
- 👤 Eduardo I. Marcelino
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 18:14
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/86
Claims Analysis
Total Claims: 29
Claim 1
Independent Claim
132 words
A method for performing an activated partial thromboplastin time test (APTT) comprising the steps of: applying non-anticoagulated blood to a test reservoir containing effective amounts of an anticoagulant, a blood coagulation activator and a platelet factor substitute to enable said applied blood to simultaneously anticoagulate and initiate contact activation, wherein said anticoagulant is a citrate compound, said coagulation activator is either diatomeceous earth, kaolin, celite, silica, micronized silica, ellagic acid or a combination thereof and said platelet factor substitute is a phospholipid. incubating the mixture at a given temperature for a given period of time and then adding a sufficient amount of calcium to said mixture to initiate the clotting cascade, measuring the length of time required to form a blood clot in said test reservoir after the addition of said calcium.
Claim 2
Independent Claim
132 words
A method for performing an activated partial thromboplastin time test (APTT) comprising the steps of: applying non-anticoagulated blood to a test reservoir containing effective amounts of an anticoagulant, a blood coagulation activator and a platelet factor substitute to enable said applied blood to simultaneously anticoagulate and initiate contact activation, wherein said anticoagulant is a citrate compound, said coagulation activator is either diatomeceous earth, kaolin, celite, silica, micronized silica, ellagic acid or a combination thereof and said platelet factor substitute is a phospholipid. incubating the mixture at a given temperature for a given period of time and then adding a sufficient amount of calcium to said mixture to initiate the clotting cascade, measuring the length of time required to form a blood clot in said test reservoir after the addition of said calcium.
Claim 3
Dependent Claim
31 words
The method according to claim 1 wherein the step of incubating includes placing the mixture in an environment having a temperature of 37° C. for a period of about three minutes.
Claim 4
Dependent Claim
13 words
The method according to claim 1, wherein said added calcium is calcium chloride.
Claim 5
Dependent Claim
12 words
The method according to claim 1 wherein said anticoagulant is sodium citrate.
Claim 6
Dependent Claim
19 words
The method according to claim 1 including the step of agitating the mixture prior to the step of incubating.
Claim 7
Dependent Claim
26 words
The method according to claim 1, wherein said added calcium is a calcium salt selected from calcium chloride, calcium bromide, D-gluconic acid calcium salt, calcium phosphate.
Claim 8
Dependent Claim
13 words
The method according to claim 1, wherein said activator is a particulate activator.
Claim 9
Dependent Claim
21 words
The method according to claim 1, wherein said activator is a soluble chemical activator, namely, tanning agents such as ellagic acid.
Claim 10
Dependent Claim
16 words
The method according to claim 1, wherein said platelet factor substitute is derived from bovine brain.
Claim 11
Dependent Claim
33 words
The method according to claim 1, wherein said platelet factor substitute is a phospholipid obtained from brain or lung tissue selected from human, bovine, ovine, goat or other animal brain or lung tissue.
Claim 12
Dependent Claim
103 words
A reagent combination used to perform an activated partial thromboplastin time (APTT) test which combination is placed in the blood containing reservoir of a blood collection device to which fresh blood is to be added, said combination comprising: a blood coagulation activator, an anticoagulant and a platelet factor substitute in effective amounts to simultaneously anticoagulate and initiate contact activation together with stabilizers and buffers, wherein said coagulation activator is a citrate compound, said platelet factor substitute is a phospholipid, said stabilizer is either sodium azide, albumin or a combination thereof, and said buffer is either sodium barbital, sodium chloride of a combination thereof.
Claim 13
Dependent Claim
103 words
A reagent combination used to perform an activated partial thromboplastin time (APTT) test which combination is placed in the blood containing reservoir of a blood collection device to which fresh blood is to be added, said combination comprising: a blood coagulation activator, an anticoagulant and a platelet factor substitute in effective amounts to simultaneously anticoagulate and initiate contact activation together with stabilizers and buffers, wherein said coagulation activator is a citrate compound, said platelet factor substitute is a phospholipid, said stabilizer is either sodium azide, albumin or a combination thereof, and said buffer is either sodium barbital, sodium chloride of a combination thereof.
Claim 14
Dependent Claim
19 words
The reagent according to claim 11, wherein said reagent is freeze dried as placed in said blood collection device.
Claim 15
Dependent Claim
15 words
The reagent according to claim 11, wherein said blood collection reservoir is a test tube.
Claim 16
Dependent Claim
12 words
The reagent according to claim 11, wherein said anticoagulant is sodium citrate.
Claim 17
Dependent Claim
19 words
The reagent according to claim 11, wherein said platelet factor substitute is a phospholipid derived from bovine brain tissue.
Claim 18
Dependent Claim
21 words
The reagent according to claim 11, wherein said stabilizers and buffers include a buffered saline solution, of sodium azide and albumin.
Claim 19
Dependent Claim
46 words
The reagent according to claim 16, wherein said buffered saline solution is barbital buffered saline comprising between 0.59 to 3.52 milligrams of sodium barbital and between 0.73 to 4.38 milligrams of sodium chloride for 1.5 cc of fresh blood to be used in said APPT test.
Claim 20
Dependent Claim
108 words
A reagent mixture used to perform an activated partial thromboplastin time (APTT) test which reagent mixture is placed in the blood containing reservoir of a blood collection device to which 1.5 cc of fresh blood is to be added comprising: between 1.5 to 15 milligrams of a diatomeceous earth as a coagulation activator, between 4.5 to 8.4 milligrams of sodium citrate as an anticoagulant and between 0.03 to 0.3 milligrams of a phospholipid as a platelet factor substitute and suitable buffers and stabilizers, wherein said buffers are either sodium barbitol, sodium chloride or a combination thereof, and said stabilizers are either sodium azide, albumin or a combination thereof.
Claim 21
Dependent Claim
108 words
A reagent mixture used to perform an activated partial thromboplastin time (APTT) test which reagent mixture is placed in the blood containing reservoir of a blood collection device to which 1.5 cc of fresh blood is to be added comprising: between 1.5 to 15 milligrams of a diatomeceous earth as a coagulation activator, between 4.5 to 8.4 milligrams of sodium citrate as an anticoagulant and between 0.03 to 0.3 milligrams of a phospholipid as a platelet factor substitute and suitable buffers and stabilizers, wherein said buffers are either sodium barbitol, sodium chloride or a combination thereof, and said stabilizers are either sodium azide, albumin or a combination thereof.
Claim 22
Dependent Claim
28 words
The reagent according to claim 18 including between 0.59 to 3.52 milligrams of sodium barbital and between 0.73 to 4.38 milligrams of sodium chloride as a buffer solution.
Claim 23
Dependent Claim
28 words
The reagent according to claim 19, further including between 0 to 3.0 milligrams of sodium azide and between 0 to 4.5 milligrams of albumin as stabilizers and preservative.
Claim 24
Dependent Claim
33 words
The reagent according to claim 20, further including adding between 1.76 to 2.94 milligrams of calcium chloride to said reagent and blood solution after a suitable incubation period to start said APPT test.
Claim 25
Independent Claim
163 words
A method for performing an activated partial thromboplastin time (APPT) test comprising the steps of: adding a predetermined amount of non-anticoagulated blood to a test reservoir containing a reagent mixture having between 1.5 to 15 milligrams of diatomaceous earth as a blood coagulation activator, between 4.5 to 8.4 milligrams of sodium citrate as an anticoagulant, between 0.03 to 0.3 milligrams of a phospholipid as a platelet factor substitute, between 0.59 to 3.52 milligrams of sodium barbital and between 0.73 to 4.38 milligrams of sodium chloride with said sodium barbital and chloride serving as a buffer, with 0 to 3.0 milligrams of sodium azide, with 0 to 4.5 milligrams of albumin, incubating said blood and reagent mixture at a temperature of 37° C. for a period between 2-5 minutes, adding between 1.76 to 2.94 milligrams of calcium chloride to said mixture after said period and, measuring the length of time required to form a blood clot in said mixture after adding said calcium chloride.
Claim 26
Independent Claim
163 words
A method for performing an activated partial thromboplastin time (APPT) test comprising the steps of: adding a predetermined amount of non-anticoagulated blood to a test reservoir containing a reagent mixture having between 1.5 to 15 milligrams of diatomaceous earth as a blood coagulation activator, between 4.5 to 8.4 milligrams of sodium citrate as an anticoagulant, between 0.03 to 0.3 milligrams of a phospholipid as a platelet factor substitute, between 0.59 to 3.52 milligrams of sodium barbital and between 0.73 to 4.38 milligrams of sodium chloride with said sodium barbital and chloride serving as a buffer, with 0 to 3.0 milligrams of sodium azide, with 0 to 4.5 milligrams of albumin, incubating said blood and reagent mixture at a temperature of 37° C. for a period between 2-5 minutes, adding between 1.76 to 2.94 milligrams of calcium chloride to said mixture after said period and, measuring the length of time required to form a blood clot in said mixture after adding said calcium chloride.
Claim 27
Dependent Claim
61 words
The method according to claim 22, wherein said predetermined amount of non-coagulated blood is 1.5 cubic centimeters of blood added to said reagent mixture having 4.5 milligrams of diatomaceous earth, 6.5 milligrams of sodium citrate, 0.075 milligrams of a phospholipid, 1.76 milligrams of sodium barbital, 2.19 milligrams of sodium chloride, with 0.75 milligrams of albumin, and 0.75 milligrams of sodium azide.
Claim 28
Dependent Claim
12 words
The method according to claim 23, wherein said albumin is bovine albumin.
Claim 29
Dependent Claim
23 words
The method according to claim 22, where the step of incubating the mixture at 37° C. is for a period of 3 minutes.
References Cited (Backward Citations)
US4782026A
Cited
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