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Method for analyzing the platelets of a blood sample

A technology of blood samples and platelets, which is applied to the analysis of materials, biomaterials analysis, material stimulation analysis, etc., can solve the problems of RNase processing insensitivity, false positives, etc.

Pending Publication Date: 2021-04-27
ESTAB FR DU SANG +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as mentioned above, some acridine oranges bind to mitochondrial nucleic acids or nucleotides in dense granules, which are not sensitive to RNase treatment, possibly producing false positives

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0125] Materials and methods

[0126] Reagent

[0127] Reagents are commercially available. Thiazole orange (TO) was from Sigma-Aldrich. Xylazine and ketamine (Imalgene ) from Bayer and Merial, respectively. Biotin was provided by ThermoFischer and diphtheria toxin (DT) was provided by Santa Cruz.

[0128] Antibodies and Flow Cytometry

[0129] In our laboratory, a rat monoclonal antibody was prepared against the extracellular domain of the platelet glycoprotein GPIbβ (Ram1, [7]) and conjugated to Alexa647 or 488. Anti-HLAI mouse antibody (clone W6 / 32) conjugated to allophycocyanin (APC) (catalog no. 311410), human anti-β2m conjugated to PE-CY7 (catalog no. 316318) and APC-cy7 (clone 2D1 ) Conjugated human anti-CD45 (cat. no. 368516) was from Biolegend. Mouse anti-H2 (clone M1 / 42) conjugated to phycoerythrin (PE) (item number: 125506) and streptavidin conjugated to APC-CY7 (item number: 405208) were purchased from Biolegend. Mouse (clone S19.8) anti-b2 microglobul...

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Abstract

The present invention relates to a method for analyzing the platelets present in a blood sample, said method comprises the steps of (a) adding to said sample a ligand which binds to the MCH I coupled to a fluorochrome, (b) measuring the mean fluorescence intensity of the platelets (MFIplatelets) with a flow cytometer and c) measuring the mean "Forward Scatter" parameter (FSC) with said flow cytometer and determining the MFIplatelets / FSC ratio. The invention also relates to an in vitro method for diagnosing a peripheral or central thrombocytopenia in a subjet.

Description

technical field [0001] The present invention relates to the field of in vitro blood analysis methods, in particular methods for the analysis of platelets present in a blood sample. More specifically, the methods described herein can be used to diagnose peripheral thrombocytopenia or central thrombocytopenia in a subject in vitro. Background technique [0002] Platelets are blood cells that lack a nucleus and are produced by the division of megakaryocytes in the bone marrow. They are usually present in human blood at an average concentration of 150,000 to 400,000 cells / μL and play an important role in hemostasis, a biological process that encompasses all phenomena that prevent and stop bleeding. [0003] Many conditions are associated with abnormalities in the number of platelets in the blood. These may be an increased concentration of platelets in the blood (called thrombocytosis), or a decreased concentration of platelets in the blood (called thrombocytopenia). [0004] ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/14G01N21/64G01N15/10G01N33/50G01N33/86G01N21/47
CPCG01N15/1012G01N15/1459G01N33/50G01N33/86G01N2015/1018G01N2800/222G01N2800/32G01N2800/50G01N2800/52G01N21/6428G01N21/6486G01N21/47G01N33/4915G01N33/582
Inventor 布兰丁·梅特凯瑟琳·安格尼厄亨利·德拉萨勒克里斯蒂安·加切特
Owner ESTAB FR DU SANG