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Method for classifying/counting leukocytes, reagent kit for classifying leukocytes, and reagent for classifying leukocytes

A technology for counting and counting leukocytes, applied in biochemical equipment and methods, analytical materials, biological testing, etc., can solve the problems of increase and decrease of blood cell count

Active Publication Date: 2013-12-18
SYSMEX CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when the subject has a disease, the number of specific blood cells increases or decreases

Method used

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  • Method for classifying/counting leukocytes, reagent kit for classifying leukocytes, and reagent for classifying leukocytes
  • Method for classifying/counting leukocytes, reagent kit for classifying leukocytes, and reagent for classifying leukocytes
  • Method for classifying/counting leukocytes, reagent kit for classifying leukocytes, and reagent for classifying leukocytes

Examples

Experimental program
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Embodiment approach

[0035] In the embodiment of the present invention, the biological sample is not particularly limited as long as it is a body fluid sample containing leukocytes. Examples of living body samples include blood, bone marrow fluid, urine, and blood component transfusions collected from mammals, preferably humans, and the like. In addition, the biological sample may be a sample that may contain abnormal leukocytes. In the present invention, leukocytes include normal leukocytes and abnormal leukocytes. Normal white blood cells are usually classified into 4 types of lymphocytes, monocytes, eosinophils, and granulocytes other than eosinophils, or are classified into lymphocytes, monocytes, and neutrophils Five types of cells, eosinophils, and, basophils.

[0036] In this specification, abnormal leukocytes generally refer to leukocytes that do not exist in peripheral blood. Examples of such abnormal leukocytes include atypical lymphocytes and blast cells. The atypical lymphocyte ref...

Embodiment 1

[0103] In this example, the concentration of the aromatic organic acid in the second reagent and the second reagent pH.

[0104] As the first reagent, Stromatolyser 4DS (Sysmex Corporation) was used. Dodecyltrimethylammonium chloride (LTAC: Tokyo Chemical Industry Co., Ltd.), polyoxyethylene (30) cetyl ether (BC30TX: Nikko Chemicals Co., Ltd.), potassium hydrogen phthalate (hereinafter referred to as phthalate Acid: Wako Pure Chemical Industries, Ltd.) and EDTA-2K (Chubu Chelest Co., Ltd.) were mixed to have the composition shown in Table 1 below to prepare a second reagent. In addition, NaOH solution was used for pH adjustment.

[0105] In addition, LTAC is a cationic surfactant, BC30TX is a nonionic surfactant, and phthalic acid is an aromatic organic acid.

[0106] 【Table 1】

[0107]

[0108] As shown in Table 1, only the second reagents A and B contained HEPES (Dojin Chemical Laboratory Co., Ltd.) as a buffer solution. The second reagents A and B correspond to the ...

Embodiment 2

[0122] In this example, using the first reagent of Example 1 and the second reagent B, G, or O, a blood test sample containing abnormally morphological lymphocytes (hereinafter referred to as abnormal test sample 1) was measured. In addition, the measurement by FCM and the calculation of the center-of-gravity distance were performed similarly to Example 1.

[0123] A scattergram (X-axis: side-scattered light intensity, Y-axis: fluorescence intensity) created from the measurement of FCM using each reagent is shown in figure 2 .

[0124] The center-of-gravity distance when the abnormal test sample 1 was measured using the second reagent B (phthalic acid concentration: 20 mM, pH 7.0) was 62.9. The center-of-gravity distance when the abnormal test sample 1 was measured using the second reagent G (phthalic acid concentration 20 mM, pH 6.0) was 69.7. The center-of-gravity distance when the abnormal test sample 1 was measured using the second reagent O (phthalic acid concentration...

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Abstract

Provided is a method for classifying / counting leukocytes, which can classify / count normal leukocytes and enables the distinction between blasts and atypical lymphocytes. The method for classifying / counting leukocytes comprises: a step of mixing a biological sample, a first reagent and a second reagent together to prepare a measurement sample, wherein the first reagent can stain a nucleic acid and the second reagent can hemolyze erythrocytes and can damage cell membranes of leukocytes to the extent that a fluorescent dye can go through the cell membranes and comprises a cationic surfactant, a nonionic surfactant and an aromatic organic acid at a concentration of 20 to 50 mM inclusive; a step of irradiating the measurement sample with light to obtain information on scattered light and information on fluorescence; and a step of classifying the leukocytes in the biological sample on the basis of the information on scattered light and the information on fluorescence obtained, and detecting blasts and atypical lymphocytes separately. In the method, the pH value of the second reagent is 5.5 to 6.4 inclusive when the concentration of the aromatic organic acid in the second reagent is 20 mM or more and less than 30 mM, and the pH value is 5.5 to 7.0 inclusive when the concentration is 30 to 50 mM inclusive.

Description

【Technical field】 [0001] The invention relates to a method for differentially counting white blood cells in a living body sample. The invention further relates to a white blood cell classification kit and a white blood cell classification reagent for classifying and counting white blood cells in living samples. 【Background technique】 [0002] Normal white blood cells are generally classified into five types: lymphocytes, monocytes, neutrophils, eosinophils, and basophils. In normal peripheral blood, each of these blood cells exists in a certain ratio. However, when the subject has a disease, the number of specific blood cells increases or decreases. Therefore, in the field of clinical examination, by performing differential counts of white blood cells, extremely useful information for disease diagnosis can be obtained. [0003] Cells that do not exist in normal peripheral blood appear in diseases such as hematopoietic tumors and viral infections. For example, in acute le...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/49G01N15/14G01N21/49G01N21/64C12Q1/02
CPCG01N21/49G01N21/6428G01N15/1429G01N15/147G01N33/5094G01N2015/011G01N2015/016
Inventor 大塚佐绪里八町和树水上利洋小国振一郎
Owner SYSMEX CORP
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