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Method for diagnosis of thrombotic disorders

a thrombotic disorder and thrombotic disease technology, applied in the field of hematology and blood coagulation disorders, can solve the problems of not including the methodology of testing the two groups of patients described above, and testing cannot be used on plasma from two important patients

Inactive Publication Date: 2005-11-24
THE SCRIPPS RES INST & RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the test cannot be used on plasma from two important groups of patients: patients taking oral anticoagulants, such as warfarin (Svensson, supra), and patients using a lupus anticoagulant (Bokarewa, et al, Blood Coagul Fibrinolysis, 5: 37, 194; Hampton, et al, N. Engl. J. Med., 331: 130, 1994).
In addition, this test does not include methodology for testing the two groups of patients described above, e.g., patients taking oral anticoagulants, such as warfarin, and patients using a lupus anticoagulant.

Method used

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  • Method for diagnosis of thrombotic disorders
  • Method for diagnosis of thrombotic disorders
  • Method for diagnosis of thrombotic disorders

Examples

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Effect test

example 1

Materials and Methods

1. Test Plasma Samples

[0040] Four and one-half ml of venous blood were collected into a siliconized BD Vacutainer tube containing 0.5 ml of 3.2% sodium citrate. Platelet poor plasma was prepared by centrifugation at 3,000 rpm for 15 minutes at room temperature. Plasma samples were kept frozen in capped plastic tubes at −70° C. to −80° C. until tested.

[0041] Plasma was obtained from the following sources: (1) 39 asymptomatic volunteers from the staff of the Laboratory Service of UCSD Medical Center and the staff of a research laboratory; (2) 21 randomly selected patients receiving oral anticoagulant therapy with warfarin; (3) 29 patients receiving oral anticoagulant activity as determined from a combination of a) a prolonged APTT that failed to correct upon a 1:1 mix with normal plasma, b) increasing apparent coagulant activity with increasing dilution of the plasma in assays for factors VIII, IX, XI, and XII, and c) an abnormal dilute Russell's viper venom t...

example 2

Stand of the Analysis of Test Results

[0047]FIG. 1 shows primary and secondary plots obtained on FV assay with and without APC of dilutions of normal pooled reference plasma. FIG. 1A shows reference curves obtained by plotting the log of clotting times against log of percent concentration of pooled normal reference plasma (primary plot) for the FV assay performed with (▴) and without (o) APC. (Y) a hypothetical clotting time obtained with a test sample which is converted to an equivalent dilution (X) of pooled normal reference plasma via steps (a) and (b). FIG. 1B shows a plot of the log of the difference in the clotting time obtained in the FV assay with and without APC against the log of percent concentration of pooled normal reference plasma. The lines in both plots are linear regression lines of the data. (♦) clotting time differences derived from data of FIG. 1A; (⋄) hypothetical result from a normal subject; (▪) hypothetical result from a heterozygous FY R506Q subject.

[0048]F...

example 3

Test Results From Control Subjects

[0053] The clotting time differences obtained on performing FV assays with and without APC on two or three dilutions of each of the plasma samples obtained from 39 control subjects are illustrated in secondary plots FIG. 2A. FIG. 2A) clotting time differences obtained when the FV assay was performed with and without APC for dilutions of plasma from control subjects are plotted against their respective equivalent dilutions of pooled normal reference plasma. (♦) pooled normal reference plasma; (⋄) 37 of the control subjects; (▪, o) two control subjects subsequently discovered to be heterozygotes for FV R506Q. Lower dashed line is regression line of the data obtained with dilutions of plasma from heterozygotes for FV R506Q. The mean regression line of the secondary plots obtained from assays of dilutions of pooled normal reference plasma on 10 different days is indicated by the solid line. The dotted lines delineate the 95% confidence limits of the sl...

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Abstract

The present invention provides an in vitro method useful for the diagnosis of a thrombotic disorder in a subject, having or at risk of having the disorder. Specifically, the disorder exemplified herein is associated with APC resistant Factor V and Va. The clotting time of a test sample is analyzed in the presence and absence of APC and compared with a standard reference sample in order to diagnose the subject.

Description

[0001] This application is a divisional of co-pending U.S. application Ser. No. 08 / 339,828, filed Nov. 14, 1994, now U.S. Pat. No. 5,705,395.[0002] This invention was made with government support by grant nos. HL 27234, HL 31950, and HL 21544 from the National Institutes of Health. The government has certain rights in the invention.BACKGROUND OF THE INVENTION [0003] 1. Field of the Invention [0004] This invention relates generally to the field of hematology and blood coagulation disorders and specifically to a method of diagnosing and screening subjects for a thromboembolic disorder. [0005] 2. Description of Related Art [0006] In 1993, Dahlback and coworkers (Dahlback, et al., Proc. Natl. Acad. Sci. USA, 90: 1004, 1993), described a previously unrecognized mechanism for familial thromboembolic disease characterized by a failure of activated protein C (APC) to appropriately prolong the activated partial thromboplastin time (APTT) of plasma. Inadequate APC-induced prolongation of the ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/56G01N33/86
CPCG01N33/86G01N2333/745G01N2333/96444G01N2405/04G01N2333/96463G01N2400/40G01N2333/96461
Inventor GRIFFIN, JOHN HRAPAPORT, SAMUEL I.LE, DZUNG T.
Owner THE SCRIPPS RES INST & RGT UNIV OF CALIFORNIA
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