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Method for measuring coagulant factor activity in whole blood

A technology of blood and factors, applied in the field of medical diagnostics and disease prevention

Inactive Publication Date: 2002-09-11
THE SCRIPPS RES INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there are no whole blood coagulability assays to accurately assess this hypercoagulable state

Method used

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  • Method for measuring coagulant factor activity in whole blood
  • Method for measuring coagulant factor activity in whole blood
  • Method for measuring coagulant factor activity in whole blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] The whole blood assay for tissue factor, described in this example, involved a protocol performed on a Hemochron instrument. This procedure used an anti-TF antibody inhibition assay to assess endogenous circulating tissue factor levels in whole blood. Materials and Reagents Required for Evaluation of TF in Circulation

[0060] Hemochron P213 Sample Tubes

[0061] 0.01M calcium chloride stock solution

[0062] Control vials containing non-inhibiting antibodies

[0063] Vial containing dried anti-tissue factor antibody

[0064] Test Quality Control Reagents

[0065] Hemoliance RecombiPlasTin stock solution (lipidated recombinant tissue factor)

[0066] TF Diluent (20mM HEPES 150mM NaCl, pH7.4, containing 0.10mg / mL

[0067] bovine serum albumin)

[0068] Blood containing anticoagulant citrate fluid and corn trypsin inhibitor (CTI)

[0069] Collection Tube Hemochron P213 Tube Preparation

[0070] Hemochron P213 tubes were preloaded with 50 μL of 0.10 M calcium chlo...

Embodiment 2

[0094] This example describes the procedure for a lipopolysaccharide-stimulation assay performed on a Hemochron instrument that evaluates the production of reactive TF in whole blood following endotoxin (lipopolysaccharide) stimulation. Materials and reagents required to evaluate reactively produced TF in whole blood following LPS stimulation

[0095] Hemochron P213 Sample Tubes

[0096] 0.10M calcium chloride stock solution

[0097] Vials containing dried LPS

[0098] Vials containing dried LPS and dried anti-tissue factor antibody

[0099] Blood containing anticoagulant citrate fluid and corn trypsin inhibitor (CTI)

[0100] Collection Tube Hemochron P213 Tube Preparation

[0101] Hemochron P213 tubes were preloaded with 50 μl of 0.10 M calcium chloride solution before the stimulated tissue factor clotting time test. Store the tubes capped at room temperature. draw blood

[0102] Discard the first few milliliters of blood drawn, then draw blood into the 5ml citrate / CTI ...

Embodiment 3

[0112] The tissue factor whole blood assay described in this example consists of a Sonoclot TM Experimental operations performed on the instrument. This procedure used an anti-TF antibody inhibition assay to assess tissue factor levels in LPS-stimulated whole blood. sample requirements

[0113] Using a 19 mm graduated needle, follow the phlebotomy procedure as detailed in, e.g., Collection, Transport, and Processing of Blood Specimens for Coagulability Testing and Coagulability Assay Procedures (National Committee on Clinical Laboratory Standards document, # H21-A-2, Volume 11, Issue 23) ((Collection.Transport and Processing of Blood Specimens for Coagulation Testing and Performance of Coagulation Assays) National Committee for Clinical Laboratory Standards document#H21-A-2, Volume XI, No. twenty three). First collect a discard tube (the top is a blue Vacutainer), and then put a plastic Vacutainer tube containing 50 μg / ml corn trypsin inhibitor (CTI) and 0.5ml 3.2% sodium c...

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Abstract

The present invention relates to the rapid assessment of the overall clotting properties of a patient's blood sample by measuring and comparing clotting times with and without inhibitors of procoagulants or anticoagulants. When the sample is whole blood, the resulting clotting time represents the total coagulation activity of the plasma and cellular components in the blood, which may be indicative of existing or impending pathology resulting from abnormal coagulation. The present invention also provides a method for determining a patient's risk of developing a thrombotic process by functionally determining the current level of one or more procoagulant or anticoagulant substances in whole blood. In addition, the present invention also provides a method for measuring the effectiveness of anticoagulant therapy and a kit for implementing the method of the present invention.

Description

[0001] This invention was supported by the National Institutes of Health (NIH) under contract number HL48872. The US Government has certain rights in this invention. technical field [0002] The present invention relates generally to the fields of medical diagnostics and disease prevention. More specifically, the present invention relates to a method for the rapid assessment of blood coagulation by determining the rate of blood clot formation using a whole blood sample in the presence and absence of at least one inhibitor of a procoagulant or an anticoagulant Active diagnostic methods and test kits. Coagulation activity in individual blood samples, as well as differences in activity between different samples, indicate the presence or potential development of certain pathological conditions. Background technique [0003] The tendency of blood to form clots too rapidly is important in anticipating the onset, development and recovery of several serious pathological states. T...

Claims

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Application Information

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IPC IPC(8): C12Q1/56G01N33/48G01N33/86
CPCC12Q1/56G01N33/86
Inventor N·麦克曼J·J·麦克唐奈
Owner THE SCRIPPS RES INST
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