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Method for diagnosing haemostasis disorders using activated charcoal

An activated carbon and barrier technology, which is applied in the fields of disease diagnosis, biochemical equipment and methods, and microbial determination/examination, and can solve problems such as the relevance of diagnosing hemostatic disorders that have not been considered.

Pending Publication Date: 2020-09-18
那慕尔大学ASBL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, the relevance of these techniques in diagnosing hemostatic disorders has not been considered

Method used

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  • Method for diagnosing haemostasis disorders using activated charcoal
  • Method for diagnosing haemostasis disorders using activated charcoal
  • Method for diagnosing haemostasis disorders using activated charcoal

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0172] Example 1: Effect of DOAC present in serum samples on aPTT and PT assays for in vitro diagnosis of hemostatic disorders

[0173] Standard tests for in vitro diagnosis of hemostatic disorders, such as in hospital laboratories, are usually performed on plasma samples obtained from patients. For the preparation of such plasma samples, the blood components of the patient's blood sample are typically separated by two centrifugation steps, both at 2500g for 15 minutes ( figure 1 ). The first centrifugation step separates the blood into solid matter (eg, red and white blood cells) (ie, the lower phase) and plasma (ie, the upper phase). Subsequently, the plasma is collected and subjected to a second centrifugation step, which pellets residual blood cells and / or platelets. The upper phase (ie platelet poor plasma) obtained by the second centrifugation step can be used for hemostasis tests. However, if the patient has been treated with a direct anticoagulant (eg, DOAC), platel...

Embodiment 2

[0175] Example 2: Methods disclosed herein eliminate the effect of DOACs on aPTT and PT assays for in vitro diagnosis of hemostatic disorders

[0176] Blood samples obtained from subjects were centrifuged at 2500g for 15 minutes to separate the blood into solid matter (eg, red and white blood cells) (ie, lower phase) and plasma (ie, upper phase).

[0177] Plasma obtained from the first (and only) centrifugation step was incubated with charcoal (10 mg / ml plasma) for 5 min, and the plasma was subsequently recovered from the charcoal by passing the plasma through a filter with 0.65 μm pores ( image 3 ). Passing the plasma through the filter was achieved by brief centrifugation. Filters with 0.65 μm pores efficiently remove activated charcoal and residual blood cells and / or platelets with a size greater than 0.65 μm from plasma with minimal interference with blood coagulation tests.

[0178] Thus, it appears that recovery of plasma from activated charcoal by passing the plasma ...

Embodiment 3

[0180] Example 3: Effect of Activated Carbon Concentration on Elimination of DOAC Effects on aPTT and PT Determinations

[0181] Plasma was obtained from blood samples obtained from subjects by one centrifugation as described in Example 2. Plasma samples were incubated with different concentrations of activated charcoal (i.e. 5 mg / ml, 10 mg / ml or 15 mg / ml) for 5 min and then recovered from the activated charcoal by passing the plasma through a filter with 0.65 μm pores and activated charcoal by brief centrifugation.

[0182] Filtered plasma samples were used for aPTT and PT determinations. The results showed that even at high concentrations of rivaroxaban (e.g., 1000 ng / ml), the effect of rivaroxaban on the determination of aPTT and PT was completely abolished ( Figure 5 a and 5b). Also, 5 mg of activated charcoal per ml is enough to achieve this effect.

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Abstract

The application provides a method for the in vitro diagnosis of a haemostasis disorder in a plasma sample obtained from a subject comprising the steps of a) contacting a plasma sample obtained from said subject with activated charcoal; b) recovering said plasma sample from said activated charcoal; and c) determining the coagulation ability of said plasma sample obtained under step (b) wherein theability of said plasma sample to coagulate is indicative of the presence, progression, or severity of a haemostasis disorder in said subject, and optionally of the nature of the haemostasis disorder.

Description

technical field [0001] The present invention is in the field of methods for diagnosing hemostatic disorders. More specifically, the invention provides methods and kits for diagnosing hemostatic disorders, eg, in patients treated with anticoagulants. [0002] Background of the invention [0003] Overall, patients with spontaneous venous thromboembolism (VTE) are at high risk of recurrence once anticoagulant therapy is discontinued. Indeed, the cumulative risk of recurrent VTE is approximately 10% at 1 year, 30% at 5 years, and 50% at 10 years. Routine and specific coagulation tests are useful screening tests to detect hemostatic disorders. Several genetic disorders (eg, activated protein C resistance, protein C and protein S deficiency, antithrombin deficiency, and elevated factor VIII) increase the risk of recurrent thromboembolism. The degree of risk depends on pre-existing conditions; therefore, testing for thrombophilic risk factors should not be used as a screening of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/86
CPCG01N33/86G01N2800/224C12Q1/56
Inventor 乔纳森·杜菲斯达明·克洛德·约瑟夫·盖尔多夫琼-米歇尔·保罗·尼古拉斯·多格纳
Owner 那慕尔大学ASBL
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