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Differential blood count reagent, kit and method of differential blood count

A white blood cell classification and reagent technology, applied in chemical instruments and methods, biochemical equipment and methods, analytical materials, etc., can solve the problems of inability to classify white blood cells into five categories, and achieve the effects of wide application range, low ambient temperature, and instrument cost saving

Active Publication Date: 2014-06-25
SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method can only divide white blood cells into three types: lymphocytes, monocytes, and granulocytes, and cannot classify white blood cells into five types.

Method used

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  • Differential blood count reagent, kit and method of differential blood count
  • Differential blood count reagent, kit and method of differential blood count
  • Differential blood count reagent, kit and method of differential blood count

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preparation example Construction

[0211] Preparation method of white blood cell differential counting reagent

[0212] The invention discloses a preparation method of a white blood cell differential counting reagent. The method comprises: dissolving the reagent components of any embodiment of the invention in water to form a single-component reagent system; or dissolving the cyanine cationic compound therein. In an organic solvent, other components are dissolved in water to become a reagent system with more than two reagent components. For the organic solvent, as long as it can fully dissolve the cyanine-type cationic compound and have a certain solubility in water, there are no other special restrictions. Common organic solvents such as methanol, ethanol, ethylene glycol, glycerin, dimethyl Sulfoxide and the like are acceptable.

[0213] Differential counting of white blood cells

[0214] Another aspect of the present invention provides a method for classifying and counting leukocytes, said method comprisin...

Embodiment 1

[0230] Reagent A for differential counting of leukocytes was prepared as follows, which consisted of the following components:

[0231] The compound of the above structural formula A 0.5ppm

[0232] Decylisoquinolinium bromide 0.4g / L

[0233] Lauryl alcohol polyoxyethylene (23) ether 1.3g / L

[0234] Sodium Benzoate 2.0g / L

[0235] Methanol 50g / L

[0236] Sodium dihydrogen phosphate 3g / L

[0237] Disodium hydrogen phosphate 4.8g / L

[0238] Add 20 μl of fresh anticoagulated blood to 1 ml of reagent A above, mix for 25 seconds while maintaining the temperature at 25°C, and then use laser flow cytometry (light source: red semiconductor laser, wavelength 640nm) to detect leukocytes. The fluorescence intensity information of the cells is measured by using the side fluorescence at a measuring angle of 90 degrees, and the side scattered light intensity information of the cells is measured by using the side scattered light at a measuring angle of 90 degrees. The result is as fi...

Embodiment 2

[0240] Reagent B for Leukocyte Differentiation was formulated as follows, consisting of the following components:

[0241] The compound of the above structural formula B 0.5ppm

[0242] Tetradecylquinolinium bromide 0.1g / L

[0243] Cetyl alcohol polyoxyethylene (25) ether 1.5g / L

[0244] Benzenesulfonic acid 2.0g / L

[0245] Tris 1.2g / L

[0246] The pH was adjusted to 7.0 with hydrochloric acid.

[0247] Take a fresh anticoagulated blood sample with high immature granulocytes (IG) (microscopic examination result IG% is 2.5%), add 20 μl of the blood sample to 1ml of reagent B above, and mix under the condition that the temperature is kept at 45°C After 15 seconds, leukocytes were detected by laser flow cytometry (light source: red semiconductor laser, wavelength 640nm). The fluorescence information of the cells is measured by using the side fluorescence at a measuring angle of 90 degrees, and the side scattered light information of the cells is measured by using the side sc...

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Abstract

The present disclosure discloses a reagent for differentiating and counting leukocytes which includes: (1) cationic cyanine compounds selected from those having the following general formulae I and II; (2) cationic surfactants selected from those having the following general formulae III, IV and / or V; (3) at least one nonionic surfactant; and (4) optionally, at least one anionic compound selected from those having one or more carboxyl or sulphonyl groups; and optionally includes alcohol compounds. Also disclosed is a kit comprising the reagent for differentiating and counting leukocytes. Further disclosed is a method for differentiating and counting leukocytes using the reagent and kit. Use of the reagent, kit and method disclosed enables the identification of leukocytes in blood samples into five subtypes in a very short time, or at least achieves the differentiation and counting of four leukocyte groupings. Moreover, immature and abnormal cells can be identified.

Description

technical field [0001] The present invention relates to blood cell classification and counting reagents and blood cell classification and counting methods, more specifically, the present invention relates to blood analysis reagents for classifying and counting white blood cells, kits containing the reagents and the use of the reagents or kits Method for differential counting of white blood cells. Background technique [0002] Human blood cells are roughly divided into three categories: red blood cells, white blood cells and platelets, among which white blood cells can be further divided into monocytes, lymphocytes and granulocytes (including eosinophils, neutrophils and basophils)5 a subcategory. Blood cells are produced in the bone marrow, differentiate from immature cells, mature, and then enter the peripheral blood to perform their respective functions. The proportions of various subtypes of white blood cells in the peripheral blood of normal healthy people are constant...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/14C12Q1/06
CPCG01N33/532C07D417/06G01N33/56972G01N33/5094G01N33/582C09B23/06C09B23/083
Inventor 雷霆徐兵匡玉吉张宝华邵建辉
Owner SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD
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