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Methods and compositions for reversing platelet aggregation

A platelet and platelet counting technology, applied in the field of platelet aggregation

Pending Publication Date: 2022-04-08
TRUVIAN SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, up to 17% of EDTA-PTCP patients also exhibit this phenomenon when blood samples are collected with citrate

Method used

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  • Methods and compositions for reversing platelet aggregation
  • Methods and compositions for reversing platelet aggregation
  • Methods and compositions for reversing platelet aggregation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] blood collection

[0132] The blood used in the following examples was obtained from a venipuncture sample or a fingertip sample. Following standard phlebotomy practice, donor venipuncture blood was collected in BD Vacutainer EDTA tubes or BD Vacutainer PST lithium heparin plasma separation tubes. Follow standard phlebotomy practices and collect donor fingertip blood in the LiHep SAFE-T-FILL TM Tube or EDTASAFE-T-FIL TM tube. All blood samples were stored at room temperature until use. Samples were analyzed within 12 hours of blood collection.

[0133] Significant platelet aggregation occurred in both venipuncture and fingertip blood collected in lithium heparin tubes. Platelet aggregation appears to occur spontaneously after blood collection. For all studies, EDTA control samples were used, where venipuncture or fingertip blood was collected into EDTA tubes. EDTA venipuncture control samples were collected into EDTA blood collection tubes from the same donor usi...

Embodiment 2

[0135] EDTA alone reverses platelet aggregation in LiHep samples

[0136] A series of anticoagulants were tested for their respective effects in reversing platelet aggregation formed in blood samples. Fresh fingertip blood samples were collected into EDTA blood collection tubes and LiHep blood collection tubes as described above. The concentrated EDTA was dried at room temperature in a microcentrifuge tube using a vacuum oven for at least one hour. Immediately after collection a 200uL fraction of the LiHep sample ("LiHep+EDTA") was passed through two separate EDTA SAFE-T-FIL TM The capillary is exposed to EDTA anticoagulant twice, resulting in a final concentration of EDTA in the blood of approximately 8.8 mM. An additional 200 uL fraction ("LiHep+TruEDTA") was added to the previously prepared vacuum-dried EDTA, resulting in a final EDTA concentration of 20 mM in blood. Thirty minutes after collection, another 200uL fraction of the LiHep sample ("LiHep+EDTA 30min") was pass...

Embodiment 3

[0140] High concentrations of EDTA negatively affect the morphology of leukocytes

[0141] Next, it was examined whether EDTA exposure would affect blood cell morphology. Specifically, fresh fingertip blood samples were collected into EDTA blood collection tubes and LiHep blood collection tubes. The concentrated EDTA was dried at room temperature in a microcentrifuge tube using a vacuum oven for at least one hour. A 200 uL fraction of the LiHep sample ("1x") was added to the previously prepared vacuum-dried EDTA, resulting in a final EDTA concentration of 20 mM in blood. An additional 200 uL fraction of the LiHep sample ("3x") was added to the previously prepared vacuum-dried EDTA, resulting in a final EDTA concentration in blood of 60 mM. All samples were pulse vortexed 3 times in 5 s bursts at the maximum vortex setting. The samples were then stained with AO. Add AO to blood samples to a final concentration of 10 ug / mL, pipette to mix and incubate at room temperature for...

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Abstract

Provided herein are methods, related compositions, and kits for solubilizing platelet clots formed in a collected blood sample. The methods, compositions and kits of the invention are useful for improving blood sample quality and blood assay accuracy in laboratory environments, especially in preventing misdiagnoses such as pseudothrombocytopenia or pseudoleukocytosis.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to U.S. Provisional Patent Application Serial No. US 62 / 845,807, filed May 9, 2019. The entire content of the aforementioned application is incorporated herein by reference in its entirety. technical field [0003] The present invention relates generally to platelet aggregation, and more particularly to methods and compositions for reversing and / or inhibiting platelet aggregation in a blood sample. Background technique [0004] Platelet aggregation is a common laboratory artifact that complicates or fails to report platelet counts. Most hematology analyzes count the number of platelets by measuring their size. Particles outside a certain size range (eg, blood cells) are ignored by the platelet channel and are not counted as platelets. Therefore, when platelets in a blood sample clump together due to artifacts, the instrument will rule out these platelet clots and generate a falsely l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P7/02G01N33/49G01N33/86
CPCA61P7/02G01N33/86G01N33/49G01N1/286G01N1/38
Inventor F·Y·李R·L·希金斯J·A·霍金斯D·C·马里努茨
Owner TRUVIAN SCI INC
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