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Blood test reagent and method

A blood detection and detection reagent technology, applied in the field of blood detection reagents, can solve the problems of long incubation time, poor membrane permeability, affecting the classification and counting of reticulocytes and mature erythrocytes, etc., to achieve improved accuracy, rapid dyeing effect, and excellent dyeing The effect of action

Active Publication Date: 2010-06-09
SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These dyes all have the disadvantage of poor membrane permeability, and require a long incubation time (several minutes to tens of minutes) for staining.
Later, a new fluorescent dye auramine O (auramine O, AuO) was developed, which can greatly shorten the staining incubation time (as low as 30 seconds), but the fluorescent dye has the problem of "orientational noise" (orientational noise) , affecting differential counts of reticulocytes and mature red blood cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] The composition and content of a blood testing reagent provided by the invention are as follows:

[0087] Cyanine dyes 10mg / L

[0088] Tris 2.42g / L

[0089] Trisodium citrate 2H 2 O 17.64g / L

[0090] SDS 4mg / L

[0091] h 2 O Amount to add to 1L

[0092] (Adjust the reagent pH to 9.0 with HCl)

[0093] Wherein the compound of general formula I used as the fluorescent dye used in the blood detection reagent has the following structure:

[0094]

[0095] A blood sample was taken, and confirmed by the new methylene blue staining method recommended by the International Committee for Standardization of Hematology (ICSH), the reticulocyte percentage was 1.60%. Take 4 μL of blood sample and mix with 1 mL of the above reagents to form a cell suspension, and incubate at 42°C for 40 seconds. The cell suspension is then passed through a flow cytometer, and the forward (0°) scattered light signal and fluorescence signal of the cells are measured to generate figure 2 In ...

Embodiment 2

[0097] The composition and content of a blood testing reagent provided by the invention are as follows:

[0098] Cyanine dyes 10mg / L

[0099] Tris 2.42g / L

[0100] Trisodium citrate 2H 2 O 17.64g / L

[0101] Decyltrimethylammonium chloride 800mg / L

[0102] h 2 O Amount to add to 1L

[0103] (Adjust the reagent pH to 9.0 with HCl)

[0104] Among them, cyanine dyes have the following structure

[0105]

[0106] A blood sample was taken, and confirmed by the new methylene blue staining method recommended by the International Committee for Standardization of Hematology (ICSH), the reticulocyte percentage was 4.10%. Take 4 μL of blood sample and mix with 1 mL of the above reagents to form a cell suspension, and incubate at 42°C for 40 seconds. The cell suspension is then passed through a flow cytometer, and the forward (0°) scattered light signal and fluorescence signal of the cells are measured to generate image 3 In the scatter diagram shown, the percentage of reticuloc...

Embodiment 3

[0108] The composition and content of a blood testing reagent provided by the invention are as follows:

[0109] Cyanine dyes 10mg / L

[0110] Tris 2.42g / L

[0111] Trisodium citrate 2H 2 O 17.64g / L

[0112] Cocamidopropyl Betaine 80mg / L

[0113] h 2 O Amount to add to 1L

[0114] (Adjust the reagent pH to 9.0 with HCl)

[0115] Among them, cyanine dyes have the following structure

[0116]

[0117] A blood sample was taken, and confirmed by the new methylene blue staining method recommended by the International Committee for Standardization of Hematology (ICSH), the reticulocyte percentage was 6.80%. Take 4 μL of blood sample and mix with 1 mL of reagent to form a cell suspension, and incubate at 42°C for 40 seconds. The cell suspension is then passed through a flow cytometer, and the forward (0°) scattered light signal and fluorescence signal of the cells are measured to generate Figure 4 As shown in the scatter diagram, the percentage of reticulocytes in the sca...

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Abstract

The invention provides a blood test reagent. The blood test reagent comprises: (1) a compound which is used as fluorochrome and has the structure represented by a general formula I, wherein n, X, R1, R2, R3, R4, R5 and Y are defined in the description; and (2) a surfactant which may be a cationic surfactant, an amphoteric surfactant or an anionic surfactant. The invention also provides a method for testing the blood, which comprises the following steps: (a) mixing a blood sample and the blood test reagent of the invention to form cell suspension; (b) detecting the scattered light and the fluorescent signal of cells; and (c) classifying and counting the cells in the blood according to the scattered light and the fluorescent signal.

Description

technical field [0001] The invention relates to a blood detection reagent and a method for using the same, more specifically, the invention relates to a blood detection reagent for classifying and counting cells in blood and a blood detection method using the reagent. Background technique [0002] Reticulocytes are transitional cells between late denucleated erythrocytes in bone marrow and fully mature erythrocytes. After reticulocytes are released from bone marrow to peripheral blood, as they mature, the RNA content in the cells gradually decreases. until it disappears completely. Therefore, the amount of intracellular RNA indicates the degree of maturation of reticulocytes. Reticulocytes are the basic test for assessing erythropoietic ability in hematological diagnosis, and are the basis for anemia diagnosis, typing, and curative effect detection. They can confirm the curative effect of bone marrow chemotherapy and transplantation, and detect the curative effect of EPO (e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/48G01N15/10C09K11/02
CPCC09K2211/1011C09K2211/1029C09K11/06C09K2211/1037G01N33/5094G01N33/80G01N33/582G01N15/1459G01N2015/1006G01N2015/1402
Inventor 匡玉吉张宝华徐兵邵建辉雷霆张丽
Owner SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD
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