Reagent for immature leukocyte analysis and reagent kit

Abstract

A reagent for analyzing immature leukocyte capable of analyzing accurately immature leukocyte such as myeloblast and immature granulocyte, and mature leukocyte, which reagent has broader allowable range of sample processing conditions than conventional ones, and a reagent kit are provided. The present invention relates to a reagent for analyzing immature leukocyte containing a surfactant for giving damage to cell membrane of red blood cell and mature leukocyte in the sample, a solubilizing agent for causing contraction to damaged blood cell, sugar, a dye for staining nucleic acid.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a flow cytometer.

FIG. 2 is a first two-dimensional distribution chart in Example 1.

FIG. 3 is a second two-dimensional distribution chart in Example 1.

FIG. 4 is a first two-dimensional distribution chart in Example 2.

FIG. 5 is a second two-dimensional distribution chart in Example 2.

FIG. 6 is a first two-dimensional distribution chart in comparison example 1.

FIG. 7 is a second two-dimensional distribution chart in comparison example 1.

FIG. 8 is a first two-dimensional distribution chart in comparison example 2.

FIG. 9 is a second two-dimensional distribution chart in comparison example 2.

FIG. 10 is a graph showing results of examples 3 through 6 and comparison example 3.

FIG. 11 is a first two-dimensional distribution chart in example 7.

FIG. 12 is a second two-dimensional distribution chart in example 7.

FIG. 13 is a first two-dimensional distribution chart in example 8.

FIG. 14 is a second two-dimensional distribution chart in example 8.

FIG. 15 is a first two-dimensional distribution chart in example 9.

FIG. 16 is a second two-dimensional distribution chart in example 9.

DESCRIPTION OF REFERENCE NUMERALS
6: Nozzle           21: Light source
22: Collimated lens 23: Flow cell
24: Collecting lens 25: Pinhole plate
26: Forward-scattered light detector
27: Collecting lens 28: Dichroic mirror
29: Side-scattered light detector
30: Pinhole plate
31: Side-fluorescence detector
32: Amplifier       33: Amplifier
34: Amplifier       35: Analyzer unit

Claims

1. A reagent for analyzing immature leukocyte contained in a sample, which reagent comprising a surfactant for giving damage to cell membrane of red blood cell and mature leukocyte, a solubilizing agent for causing contraction to damaged blood cell, a sugar, and a dye for staining nucleic acid.

2. The reagent according to claim 1, wherein said surfactant is a polyoxyethylene-type nonionic surfactant represented by the following chemical formula; R1—R2—(CH2CH2O)n—H

3. The reagent according to claim 1 wherein said solubilizing agent is at least one selected from the group consisting of sarcosine derivative, salt of sarcosine derivative, cholic acid derivative, and methylglucamide.

4. The reagent according to claims 1, wherein said sugar is at least one selected from the group consisting of xylitol, arabinose, glucose, mannitol, sorbitol, ribitol.

5. The reagent according to claims 1 containing from 10 to 75 g/L of said sugar.

6. The reagent according to claims 1, which does not substantially contain sodium chloride.

7. The reagent according to claims 1, pH of which is from 5.0 to 9.0.

8. The reagent according to claims 1, osmotic pressure of which is from 150 to 600 mOsm/kg.

9. A reagent kit for analyzing immature leukocyte contained in a sample comprising a first reagent which comprises a surfactant for giving damage to cell membrane of red blood cell and mature leukocyte, a solubilizing agent for causing contraction to damaged blood cell, and an osmotic pressure regulator, having osmotic pressure of from 150 to 600 mOsm/kg and electric conductivity of less than 6 mS/cm, and a second reagent containing a dye for staining nucleic acid.

10. The reagent kit according to claim 9, wherein the first reagent has electric conductivity of less than 3 mS/cm.

11. The reagent kit according to claim 9, wherein said osmotic pressure regulator is at least one selected from the group consisting of sugar and amino acid.

12. The reagent kit according to claim 11, wherein said amino acid is at least one selected from the group consisting of glycine and alanine.

13. The reagent according to claim 11 containing from 1 to 50 g/L of said amino acid.

14. The reagent according to claims 9, wherein concentration of sodium chloride of said first reagent is less than 3 g/L.

15. A reagent kit for analyzing immature leukocyte contained in a sample comprising a first reagent which comprises a surfactant for giving damage to cell membrane of red blood cell and mature leukocyte, a solubilizing agent for causing contraction to damaged blood cell, sugar, a second reagent containing a dye for staining nucleic acid.

16. The reagent according to claim 15, wherein said surfactant is a polyoxyethylene-type nonionic surfactant represented by the following chemical formula; R1—R2—(CH2CH2O)n—H

17. The reagent according to claim 15 wherein said solubilizing agent is at least one selected from the group consisting of sarcosine derivative, salt of sarcosine derivative, cholic acid derivative, and methylglucamide.

18. The reagent according to claims 15, wherein said sugar is at least one selected from the group consisting of xylitol, arabinose, glucose, mannitol, sorbitol, ribitol.

19. The reagent according to claims 15 containing from 10 to 75 g/L of said sugar.

20. The reagent according to claims 15, which does not substantially contain sodium chloride.