Claims
- 1. A diagnostic prothrombin time reagent comprising an extracted thromboplastin composition said prothrombin time reagent having approximately normal prothrombin times and a sensitivity to blood coagulation factor VII deficiency of at least forty seconds at a concentration of about 10% of said extracted thromboplastin composition and sensitivity to coumadin therapy greater than 1.8, as measured by the ratio of the prothrombin time value of an abnormal sample to a normal sample and wherein the extracted thromboplastin composition is made by the process comprising:
- (a) contacting rabbit brain tissue containing thromboplastin with an effective amount of extraction fluid comprising about 0.1 to 1.0 grams of barium sulfate per gram of tissue, about 0.01 to 0.25% of a nonionic detergent, about 5 to 100 mM chaotropic ion, and a salt, thereby extracting the thromboplastin into the fluid, and
- (b) separating said extracted thromboplastin from said depleted tissue and barium sulfate.
- 2. The diagnostic prothrombin time reagent of claim 1 wherein the amount of extraction fluid is about 100 mL for every 5 grams of tissue.
- 3. The diagnostic prothrombin time reagent of claim 1 wherein the concentration of the salt is from about 30 to 180 mM.
- 4. The diagnostic prothrombin time reagent of claim 1 wherein the normal prothrombin time ranges from about 9 to 15 seconds and the enhanced sensitivity to coumadin therapy shown by COL2/COL1 ratio ranges from about 1.8 to 3.0.
- 5. A diagnostic prothrombin time reagent comprising an extracted thromboplastin composition said prothrombin time reagent having approximately normal prothrombin times and sensitivity to blood coagulation factor VII deficiency of at least sixty-five seconds at a concentration of about 32% of said extracted thromboplastin composition and sensitivity to coumadin therapy greater than 1.8, as measured by the ratio of the prothrombin time value of an abnormal sample to a normal sample and wherein the extracted thromboplastin composition is made by the process comprising:
- (a) contacting rabbit brain tissue containing thromboplastin with an effective amount of extraction fluid comprising about 5 to 100 mM chaotropic ions, thereby extracting the thromboplastin into the fluid, and
- (b) separating said extracted thromboplastin from said depleted tissue.
- 6. The diagnostic prothrombin time reagent of claim 5 wherein the amount of extraction fluid is about 100 mL for every 5 grams of tissue.
- 7. The diagnostic prothrombin time reagent of claim 5 wherein the normal prothrombin time ranges from about 9 to 15 seconds and the enhanced sensitivity to coumadin therapy shown by COL2/COL1 ratio ranges from about 1.8 to 3.0.
- 8. A diagnostic prothrombin time reagent comprising an extracted thromboplastin composition said prothrombin time reagent having approximately normal prothrombin times and sensitivity to blood coagulation factor VII deficiency of at least one hundred seconds at a concentration of about 32% of said extracted thromboplastin composition and sensitivity to coumadin therapy greater than 1.8, as measured by the ratio of the prothrombin time value of an abnormal sample to a normal sample and wherein the extracted thromboplastin composition is made by the process comprising:
- (a) contacting rabbit brain tissue containing thromboplastin with an effective amount of extraction fluid comprising about 0.1 to 1.0 grams of barium sulfate per gram of tissue, about 0.01 to 0.25% nonionic detergents, and a salt, thereby extracting the thromboplastin into the fluid, and
- (b) separating said extracted thromboplastin from said depleted tissue and barium sulfate.
- 9. The diagnostic prothrombin time reagent of claim 8 wherein the amount of extraction fluid is about 100 mL for every 5 grams of tissue.
- 10. The diagnostic prothrombin time reagent of claim 8 wherein the concentration of the salt is from about 30 to 180 mM.
- 11. The diagnostic prothrombin time reagent of claim 8 wherein the normal prothrombin time ranges from about 9 to 15 seconds and the enhanced sensitivity to coumadin therapy shown by COL2/COL1 ratio ranges from about 1.8 to 3.0.
Parent Case Info
This is a continuation of application Ser. No. 07/632,894, filed on Dec. 24, 1990, now abandoned, which is a division of U.S. Ser. No. 07/276,083 filed Nov. 23, 1988, now abandoned.
US Referenced Citations (1)
Number |
Name |
Date |
Kind |
5017556 |
O'Brien et al. |
May 2191 |
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Non-Patent Literature Citations (7)
Entry |
B. A. Bradlow et al. "A Human Brain Thromboplastin Standard for Distribution in South Africa" S. Africa Med. J. 50(50) 1989-1992 (1976). |
K. J. Stevenson et al, "The British Comparative Thromboplastin: . . . " Br. J. Haematology 44(3) 495-501 (1980). |
A. J. Quick "Thromboplastin as a Reagent" Thrombosis et Diathesis Haemorrhagica 23(3) 585-592 (1920). |
H. Starr et al, "Prothrombin Times: An Evaluation of Four Thromboplastins and Four Machines" Pathology 12(4) 567-574 (1980). |
Guha et al. (1986) Proc. Natl. Acad. Sci. U.S.A. 83:299-302. |
Bach et al. (1981) J. Biol. Chem. 256(16):8324-8331. |
Bjorklid et al. (1977) Biochem. J. 165:89-96. |
Divisions (1)
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Number |
Date |
Country |
Parent |
276083 |
Nov 1988 |
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Continuations (1)
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Number |
Date |
Country |
Parent |
632894 |
Dec 1990 |
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